agilent 1200 series analytical and preparative scale ... · pdf filea agilent 1200 series...

Agilent 1200 Series Analytical and Preparative ScaleFraction Collectors

User Manual

A

Agilent 1200 Series Fraction Collectors User Manual

Notices© Agilent Technologies, Inc. 2006

No part of this manual may be reproduced in any form or by any means (including elec-tronic storage and retrieval or translation into a foreign language) without prior agree-ment and written consent from Agilent Technologies, Inc. as governed by United States and international copyright laws.

Microsoft ® is a U.S. registered trademark of Microsoft Corporation.

Manual Part NumberG1364-90010

Edition02/06

Printed in Germany

Agilent TechnologiesHewlett-Packard-Strasse 8 76337 Waldbronn

Warranty

The material contained in this docu-ment is provided “as is,” and is sub-ject to being changed, without notice, in future editions. Further, to the max-imum extent permitted by applicable law, Agilent disclaims all warranties, either express or implied, with regard to this manual and any information contained herein, including but not limited to the implied warranties of merchantability and fitness for a par-ticular purpose. Agilent shall not be liable for errors or for incidental or consequential damages in connec-tion with the furnishing, use, or per-formance of this document or of any information contained herein. Should Agilent and the user have a separate written agreement with warranty terms covering the material in this document that conflict with these terms, the warranty terms in the sep-arate agreement shall control.

Technology Licenses The hardware and/or software described in this document are furnished under a license and may be used or copied only in accor-dance with the terms of such license.

Restricted Rights LegendIf software is for use in the performance of a U.S. Government prime contract or subcon-tract, Software is delivered and licensed as “Commercial computer software” as defined in DFAR 252.227-7014 (June 1995), or as a “commercial item” as defined in FAR 2.101(a) or as “Restricted computer soft-ware” as defined in FAR 52.227-19 (June 1987) or any equivalent agency regulation or contract clause. Use, duplication or disclo-sure of Software is subject to Agilent Tech-nologies’ standard commercial license terms, and non-DOD Departments and Agencies of the U.S. Government will

receive no greater than Restricted Rights as defined in FAR 52.227-19(c)(1-2) (June 1987). U.S. Government users will receive no greater than Limited Rights as defined in FAR 52.227-14 (June 1987) or DFAR 252.227-7015 (b)(2) (November 1995), as applicable in any technical data.

Safety Notices

CAUTION

A CAUTION notice denotes a haz-ard. It calls attention to an operat-ing procedure, practice, or the like that, if not correctly performed or adhered to, could result in damage to the product or loss of important data. Do not proceed beyond a CAUTION notice until the indicated conditions are fully understood and met.

WARNING

A WARNING notice denotes a hazard. It calls attention to an operating procedure, practice, or the like that, if not correctly per-formed or adhered to, could result in personal injury or death. Do not proceed beyond a WARNING notice until the indicated condi-tions are fully understood and met.

In This Guide…This manual contains technical reference information about the Agilent 1200 Series fraction collectors analytical and preparative scale. The manual describes the following:

1 Installing the Fraction Collector

This chapter provides information about the installation of the Agilent 1200 Series fraction collectors AS and PS.

2 Configuration and Operation of the Fraction Collector

This chapter describes the configuration and operation of the fraction collector including guidelines to optimize the system and to avoid problems.

3 Control Module Screens of the Fraction Collector

The reference to the screens of the Agilent 1200 Series control module is provided in the chapter.

4 Troubleshooting and Test Functions

This chapter describes the modules built-in troubleshooting and test functions.

5 Repairing the Fraction Collector

This chapter contains instructions on simple repair and maintenance procedures.

6 Parts and Materials

This chapter contains lists for identification of common repair and maintenance parts.

7 Specifications

This chapter contains performance specifications of the fraction collectors.

A Safety Information

This appendix provides a safety summary.

Agilent 1200 Series Fraction Collectors User Manual 3

4 Agilent 1200 Series Fraction Collectors User Manual

Contents

1 Installing the Fraction Collector 11

Site Requirements 12

Power Consideration 12Power Cords 13Bench Space 13Environment 14

Unpacking the Fraction Collector 16

Damaged Packaging 16Delivery Checklist 16Accessory Kits 17

Optimizing the Stack Configuration 20

Installing the Fraction Collector 25

Installing a Thermostatted Fraction Collector 29

Flow Connections to the Fraction Collector 34

Fraction Collector Trays 37

Installing the Fraction Collector Trays 37Numbering of Vial, Test Tube and Well-plate Positions 38

Configure Well-plate Types 40

Transporting the Fraction Collector 43

2 Configuration and Operation of the Fraction Collector 45

Configuration of the Fraction Collector 46

Configuration of the fraction collector in the ChemStation 46Configuring multiple fraction collectors 50


Contents

Delay volumes and delay calibration 52

Delay times and volumes 52Detector signal delay 53Performing a delay calibration with an UV detector 57Performing a delay calibration with an MSD 61Making Your Own Calibration Method 66

Setting up a Fraction Collector Method 67

Fraction Trigger Mode 68Fraction Preview 72

Starting your run with fraction collection 74

Assignment of Start Location for fraction collection 74Online tick marks 77

Viewing your Results 78

Data Analysis 78Report 79

Special Applications 80

Pooling 80Sample Recovery 81Semi-preparative operation 82Using high test tubes in the analytical scale fraction collector 83

Optimizing fraction collection 84

Limitations and how to avoid problems 85

Check-out Procedures 86

Check-out procedure for UV-triggered fraction collection AS 86Check-out procedure for UV-triggered fraction collection PS 89

Application Notes 91

3 Control Module Screens of the Fraction Collector 93

Major keys on the Agilent 1200 Series control module 94


Contents

Screens available from the Analysis view 95

Screens available from the System view 109

4 Troubleshooting and Test Functions 127

Status Indicators 128Error Messages 128Maintenance Functions 128Transport Unit Self Alignment 128

Status Indicators 129

Power Supply Indicator 129Instrument Status Indicator 130

Maintenance Functions 131

Transport Unit Self Alignment 133

Step Commands 135

Troubleshooting 136

5 Repairing the Fraction Collector 137

Introduction into Repairing the Fraction Collector 138

Simple Repairs 138Cleaning the Fraction Collector 138

Overview of Main Repair Procedures 139

Simple Repairs 140

Replacing the Inlet/Waste Tubings 141Replacing the Valve to Needle Tubing 145Exchanging the Preparative Needle Assembly 148Exchanging the Analytical Needle Assembly 150Exchanging the Diverter Valve 153Exchanging the Internal Tray 156


Contents

6 Parts and Materials 161

Supported Trays for Fraction Collectors 162

List of Recommended Test Tubes 164

List of Recommended Vials and Caps 165

List of Recommended Plates and Closing Mats 168

Transport Unit Assembly (Preparative Scale) 170

Transport Unit Assembly (Analytical Scale) 171

Needle Assemblies 173

Diverter-Valve Assembly 174

Tubing Kits 175

Internal Tray Assembly 176

Fraction Collector Accessory Kit 177

7 Specifications 179

Performance Specifications for the Fraction Collectors 180

A Safety Information 187

Safety Information 188

General 188Operation 189Safety Symbols 190

Lithium Batteries Information 191

Radio Interference 192

Test and Measurement 192

Sound Emission 193

Manufacturer’s Declaration 193


Contents

Solvent Information 194

Solvents 194

Agilent Technologies on Internet 195

Index 197


Contents


Agilent 1200 Series Fraction CollectorsUser Manual

1Installing the Fraction Collector

Site Requirements 12

Unpacking the Fraction Collector 16

Optimizing the Stack Configuration 20

Installing the Fraction Collector 25

Installing a Thermostatted Fraction Collector 29

Flow Connections to the Fraction Collector 34

Fraction Collector Trays 37

Configure Well-plate Types 40

Transporting the Fraction Collector 43

11Agilent Technologies

1 Installing the Fraction CollectorSite Requirements

Site Requirements

A suitable site environment is important to ensure optimum performance of the fraction collector.

Power Consideration

The fraction collector power supply has wide-ranging capability (see Table 1 on page 14). Consequently there is no voltage selector in the rear of the fraction collector. There are also no externally accessible fuses, as automatic electronic fuses are integrated in the power supply.

The thermostatted fraction collector comprises two modules, the fraction collector (G1364B or G1364C) and the thermostat (G1330A). Both modules have a separate power supply and a power plug for the line connections. The two modules are connected by a control cable and both are turned on by the fraction collector module. The thermostat power supply has two externally accessible fuses.

WARNING To disconnect the fraction collector from line power, unplug the power cord. The power supply still uses some power, even if the power switch on the front panel is turned off.

WARNING To disconnect the thermostatted fraction collector from line power, unplug the power cord from the fraction collector and the thermostat. The power supplies still use some power, even if the power switch on the front panel is turned off. Please make sure that it is always possible to access the power plug.

WARNING Shock hazard or damage of your instrumentation can result if the devices are connected to a line voltage higher than specified.


Installing the Fraction Collector 1Site Requirements

Power Cords

Your fraction collector is delivered with a power cord which matches the wall socket of your particular country or region. The plug on the power cord which connects to the rear of the instrument is identical for all types of power cord.

Bench Space

The fraction collector dimensions and weight (see Table 1 on page 14) allow the instrument to be placed on almost any laboratory bench. The instrument requires an additional 2.5 cm (1.0 inch) of space on either side, and approximately 8 cm (3.1 inches) at the rear for the circulation of air, and room for electrical connections. Ensure the fraction collector is installed in a horizontal position.

The thermostatted fraction collector dimensions and weight (see Table 2 on page 15) allow the instrument to be placed on almost any laboratory bench. The instrument requires an additional 25 cm (10 inches) of space on either side for the circulation of air, and approximately 8 cm (3.1 inches) at the rear for electrical connections. Ensure the fraction collector is installed in a horizontal position.

If a complete Agilent 1200 Series LC system is to be installed on the bench, make sure that the bench is designed to carry the weight of all the modules. For a complete system including the thermostatted fraction collector it is recommended to position the modules in two stacks, see “Recommended Stack Configuration - Fraction Collector in an Analytical System (Front View)” on page 21. Make sure that in this configuration there is 25 cm (10 inches) space on either side of the thermostatted fraction collector for the circulation of air.

WARNING Never operate your instrumentation from a power outlet that has no ground connection. Never use a power cord other than the power cord designed for your region.

WARNING Never use cables other than the ones supplied by Agilent Technologies to ensure proper functionality and compliance with safety or EMC regulations.


1 Installing the Fraction CollectorSite Requirements

Environment

Your fraction collector will work within specifications at ambient temperatures and relative humidity as described in Table 1 and Table 2 on page 15.

CAUTION Do not store, ship or use your fraction collector under conditions where temperature fluctuations may cause condensation within the fraction collector. Condensation will damage the system electronics. If your fraction collector was shipped in cold weather, leave it in its box, and allow it to warm up slowly to room temperature to avoid condensation.

Table 1 Physical Specifications - Fraction Collectors (G1364B, G1364C)

Type Specification Comments

Weight 13.5 kg (29.8 lbs)

Dimensions(height × width × depth)

200 × 345 × 440 mm(8 × 13.5 × 17 inches)

Line voltage 100 – 240 VAC, ±10 % Wide-ranging capability

Line frequency 50 or 60 Hz, ±5 %

Power consumption (apparent power) 200 VA Maximum

Power consumption (active power) 180 W Maximum

Ambient operating temperature 4 – 55 °C (41 – 131 °F)

Ambient non-operating temperature -40 – 70 °C (-4 – 158 °F)

Humidity < 95 %, at 25 – 40 °C (77 – 104 °F) Non-condensing

Operating Altitude Up to 2000 m (6500 ft)

Non-operating altitude Up to 4600 m (14950 ft) For storing the fraction collector

Safety standards: IEC, CSA, UL Installation Category II, Pollution Degree 2.For indoor use only.


Installing the Fraction Collector 1Site Requirements

Table 2 Physical Specifications - Thermostat (G1330B)

Type Specification Comments

Weight 18.5 kg (40.7 lbs)

Dimensions(height × width × depth)

140 × 345 × 435 mm(5.5 × 13.5 × 17 inches)

Line voltage 100 – 120 or 220 – 240 VAC, ± 10 % Wide-ranging capability

Line frequency 50 or 60 Hz, ± 5 %

Power consumption (active power) 210 W Maximum

Power consumption (apparent power) 260 VA Maximum

Ambient operating temperature 4 – 40 °C (41 – 131 °F)

Ambient non-operating temperature -40 – 70 °C (-4 – 158 °F)

Humidity < 95 %, at 25 – 40 °C (77 – 104 °F) Non-condensing

Operating Altitude Up to 2000 m (6500 ft)

Non-operating altitude Up to 4600 m (14950 ft) For storing the fraction collector

Safety standards: IEC, CSA, UL, EN Installation Category II, Pollution Degree 2.For indoor use only.


1 Installing the Fraction CollectorUnpacking the Fraction Collector

Unpacking the Fraction Collector

Damaged Packaging

Upon receipt of your fraction collector, inspect the shipping containers for any signs of damage. If the containers or cushioning material are damaged, keep them until the contents have been checked for completeness and the fraction collector has been mechanically and electrically checked. If the shipping container or cushioning material is damaged, notify the carrier and keep the shipping material for the carrier‘s inspection.

Delivery Checklist

Ensure all parts and materials have been delivered with the fraction collector. For this compare the shipment content with the checklist included in each instrument box. Please report missing or damaged parts to your local Agilent Technologies sales and service office.

Three models of Agilent 1200 Series fraction collector are available:

• G1364B Fraction Collector, preparative scale, designed for flow rates up to 100 ml / min. and for the use with vials, deep well plates and test tubes up to 100 mm height.

• G1364C Fraction Collector, analytical scale, designed for flow rates below 10 ml / min. (can be modified for higher flow rates) and for the use with vials, test tubes of up to 75 mm height, well-plates and a 40-funnel tray connecting to external locations of any size

CAUTION If you need to ship the fraction collector at a later date, always use the shipping protection foam parts (see “Transporting the Fraction Collector” on page 43).

CAUTION If there are signs of damage to the fraction collector, please do not attempt to install the fraction collector.


Installing the Fraction Collector 1Unpacking the Fraction Collector

• G1364C Thermostatted Fraction Collector, analytical scale, can be created by additionally ordering and installing a G1330B Fraction Collector Thermostat.

The terms (thermostatted) analytical scale fraction collector and the preparative scale fraction collector refer to these Agilent 1200 Series modules in this manual. Unless otherwise stated all information in this section is valid for all models.

Accessory Kits

Each shipment contents an Accessory kit with the necessary tools to install the system and to have an operating system.

Table 3 Delivery Checklist for the G1364B (preparative scale) and G1364C (analytical scale) Fraction Collector

Description Quantity

Fraction collector module with inlet / waste tubing assembly 1

Power cord, local (Matching the sockets in your country or region) 1

Accessory kit (see below) 1

Delay Calibrant G1946-85020 1

Table 4 Delivery Checklist for the G1330B Thermostat Module (optional)

Description Quantity

Thermostat Module 1

Power cord, local (Matching the sockets in your country or region) 1

Accessory kit thermostat module (see below) 1


1 Installing the Fraction CollectorUnpacking the Fraction Collector

• The Accessory kit (G1330-68705) shown in Table 6 is shipped with the (G1330B) thermostat module if the thermostat module was ordered.

Table 5 Fraction Collector Accessory Kit Contents G1364-68725

Description Quantity Part Number

Wrench, open end, 4mm 1 8710-1534

Wrench, open end, 1/4-5/16 inch 1 8710-0510

Hex key 2.0 mm 1 8710-2476

Finger tight fittings, 1/16” f-120*

* Reorder gives pack of 2

3 0100-1516

Waste tubing (1.2 m)†

† Reorder gives 5 m

1 5062-2463

Y-Type Fitting for waste tubing‡

‡ Reorder gives pack of 10

1 5065-9971

CAN cable, 1 m 1 5181-1519

Air channel adapter 1 G1329-43200

Sticking clamp for corrugated waste tubing (large)**

** Reorder gives 10 clamps for corrugated waste tubing (large))

3 5065-9976

Sticking clamp for waste tubing (small)††

†† Reorder gives 10 clamps for waste tubing (small)

3 5065-9976

ESD wrist strap 1 9300-1408

Tray for 15 x 6 ml vials 1 G1313-44513


Table 6 (Optional) Thermostat Module Accessory Kit Contents G1330-68705


Waste tubing (1.2 m)* 1 5062-2463

Cable thermostat module to fraction collector 1 G1330-81600


Installing the Fraction Collector 1Unpacking the Fraction Collector

* Reorder gives 5 m


1 Installing the Fraction CollectorOptimizing the Stack Configuration

Optimizing the Stack Configuration

If your fraction collector is part of a system, you can ensure optimum performance and minimum delay volume by installing the following configuration.

• Figure 1 and Figure 2 on page 22 show the configuration recommended for the fraction collector in an analytical scale system.

• Figure 3 on page 23 and Figure 4 on page 24 show the configuration recommended for the fraction collector with a preparative scale system.

NOTE Agilent 1200 Series LC purification systems can comprise many different LC modules. This section describes some common stack configurations and general considerations. On more complex systems, e.g. with two or more fraction collectors or with mass selective detector, other stack configurations might show advantages.


Installing the Fraction Collector 1Optimizing the Stack Configuration

Figure 1 Recommended Stack Configuration - Fraction Collector in an Analytical Sys-tem (Front View)

UV-Vis

FractionCollector

Detector

SolventCompartment

Degasser

Pump

Autosampler

ColumnCompartment

NOTE For information about connecting an Agilent LC-MSD, a Non-1200 or Non-UV-Vis detector to the system, please refer to the “User’s Guide for Purification / High Throughput System”, PN G2262-90010 or to the “Installation Note for the Mass Based Fraction Collection Kit”.



Figure 2 Recommended Stack Configuration - Fraction Collector in an Analytical Sys-tem (Rear View)

SolventCompartment

Degasser

Pump

Autosampler

ColumnCompartment

UV-Vis

FractionCollector

Detector

GPIB or LANto LC ChemStation

CAN Bus cable

NOTE If a G1330B ALS thermostat is part of the system it must always be installed underneath the thermostatted injector or fraction collector (see “Installing a Thermostatted Fraction Collector” on page 29).

NOTE The fraction collector should never be positioned on top of a module that generates heat. This could lead to an unwanted evaporation of fractions in the fraction collector (e.g. Agilent 1200 Series thermostatted column compartment G1316A/B or Agilent 1200 Series diode-array detectors G1315B/C.


Installing the Fraction Collector 1Optimizing the Stack Configuration

Figure 3 Recommended Stack Configuration - Fraction Collector in a Preparative Sys-tem (Front View)

UV-Vis

FractionCollector

Column Organizer

PreparativePump A

PreparativePump B

SolventCompartment

Autosampler

Detector

NOTE For information about connecting an Agilent LC-MSD, a Non-1200 or Non-UV-Vis detector to the system, please refer to the “User’s Guide for Purification / High Throughput System”, PN G2262-90010 or to the “Installation Note for the Mass Based Fraction Collection Kit”.



Figure 4 Recommended Stack Configuration - Fraction Collector in a Preparative Sys-tem (Rear View)

UV-Vis

FractionCollector

Detector

PreparativePump A

PreparativePump B

SolventCompartment

Autosampler

Column Organizer

GPIB or LANto ChemStation

CAN BusCable

NOTE If a G1330B ALS thermostat is part of the system it must always be installed underneath the module, that is thermostatted injector or fraction collector (see “Installing a Thermostatted Fraction Collector” on page 29).

NOTE The fraction collector should never be positioned on top of a module that generates heat, which can lead to an unwanted evaporation of fractions in the fraction collector (e.g. Agilent 1200 Series thermostatted column compartment G1316A/B or Agilent 1200 Series diode-array detectors G1315B/C.


Installing the Fraction Collector 1Installing the Fraction Collector

Installing the Fraction Collector

1 Install the LAN interface board in the fraction collector (if required).

2 Remove the adhesive tape which covers the side and front doors.

3 Open the front door and remove the left side door.

4 Remove the transport protection foam.

5 Install the corrugated waste tube in the plastic port at the front bottom center of the instrument. Slide the waste tubing coming from the internal tray (if present) through the plastic port and the corrugated waste tube (see Figure 5 on page 26). Route the corrugated waste tubing into a waste container.

Preparation Locate bench spaceProvide power connectionsUnpack the fraction collector

Parts required Fraction CollectorPower cord, Chemstation and/or Control Module G1323B

WARNING When opening capillary or tube fittings solvents may leak out. Please observe appropriate safety procedures (for example, goggles, safety gloves and protective clothing) as described in the material handling and safety data sheet supplied by the solvent vendor, especially when toxic or hazardous solvents are used.

WARNING To avoid personal injury, keep fingers away from the needle area during fraction collector operation. Do not attempt to insert or remove a vial or a plate when the needle is positioned.


1 Installing the Fraction CollectorInstalling the Fraction Collector

6 Re-install the left side door (take care of the magnet at the back). Ensure the side door is correctly installed (its presence is sensed by a hall sensor, a missing side door will result in a NOT-READY state of the instrument).

7 Place the fraction collector in the stack or on the bench in all horizontal position.

8 Ensure the power switch at the front of the fraction collector is OFF.

9 Connect the power cable to the power connector at the rear of the fraction collector.

10 Connect the CAN cable to the other Agilent 1200 Series LC modules.

11 If an Agilent ChemStation is the controller, it must be connect to the LAN interface (should be installed to the detector).

12 Connect the APG remote cable (optional) for non Agilent 1200 Series instruments.

13 Install the tray that has been delivered for your fraction collector. The test tube trays can be adjusted in height, depending on the height of the used test tubes. To adjust the height of the trays, press against the snappers at

Figure 5 Installing the Corrugated Waste Tubing in the Plastic Port

2

1


Installing the Fraction Collector 1Installing the Fraction Collector

the guides in the 4 corners of a tray and move the top plate of the tray up or down (see Figure 6 on page 27).

14 Turn ON power by pushing the button at the lower left hand side of the fraction collector.

15 The exhaust fan will turn ON and remove potential solvent vapor from the inside of the instrument. After 2 minutes close the front door. Then the fraction collector will start the hardware initialization process. At the end of this process the status LED should be green.

NOTE The preparative scale fraction collector (G1364B) and the analytical scale fraction collector (G1364C) are shipped with two trays (40x2 ml half tray and 15 x 6ml half tray).

Figure 6 Adjusting the Height of the (Full) Test Tube Trays.


1 Installing the Fraction CollectorInstalling the Fraction Collector

Figure 7 Cable Connections

RS232C

CAN-bus

Remote

CAN cable to next module

CAN cable to previous module

Control of Thermostat

CAN-DC-Out

AC-Power

NOTE The fraction collector is turned ON when the line power switch is pressed and the green indicator lamp is illuminated. The detector is turned OFF when the line power switch is protruding and the green light is OFF.

WARNING To disconnect the fraction collector from the line, unplug the power cord. The power supply still uses some power, even if the switch at the front panel is turned OFF.


Installing the Fraction Collector 1Installing a Thermostatted Fraction Collector

Installing a Thermostatted Fraction Collector

1 Place the thermostat on the bench.

2 Remove the front cover and route the condensation drain tube to a waste container.

3 Install the LAN interface board in the fraction collector (if required).

4 Remove the adhesive tape which covers the side and front doors.

5 Open the front door and remove the left side door.

6 Remove the transport protection foam.

7 Install the corrugated waste tube in the plastic port at the front bottom center of the fraction collector and route down into a waste container. Slide the waste tubing coming from the internal tray (if present) through the

Preparation Locate bench spaceProvide power connectionsUnpack the fraction collector and the thermostat

Parts required Fraction Collector and thermostatPower cords,ChemStation and/or Control Module G1323B

WARNING Make sure the condensation drain tube runs down into a waste container without any (upwards) bends or curves. Free and unrestricted flow of the condensation into a waste container must be guaranteed. Make sure that the condensation drain tube is always above the liquid level in the container. If the tube is located in liquid the condensed water cannot flow out of the tube and the outlet is blocked. Any further condensation will then remain in the instrument. This may damage the instruments electronics.

CAUTION The fraction collector thermostat requires 25 cm (10 inch) space on each for sufficient air circulation.


1 Installing the Fraction CollectorInstalling a Thermostatted Fraction Collector

plastic port and the corrugated waste tube (see Figure 5 on page 26). Route the corrugated waste tubing into a waste container.

8 Re-install the left side door (take care of the magnet at the back). Ensure the side door is correctly installed (its presence is sensed by a hall sensor, a missing side door will result in a NOT-READY state of the instrument).

9 Place the fraction collector on top of the thermostat. Make sure that the fraction collector is correctly engaged in the thermostat locks.

10 Remove the plastic cover from the tray base, place the air channel adapter (1) into the fraction collector tray base. Make sure the adapter is fully pressed down. This assures that the cold airstream from the thermostat is correctly guided to the tray area of the fraction collector. Place the plug channel (2) on top of the air channel adapter. Both devices must be installed correctly, to assure proper operation of the instrument.

11 The analytical scale fraction collector is delivered with a pre-installed tray compartment divider. This divider should only be used with the thermostatted fraction collector, if a (half or std.) tray is installed into the left and center positions of the instrument. If any tray is installed to the right side of the instrument, remove the tray compartment divider. The tray compartment divider optimizes the cooling performance of the instrument, if only the left and center position of the fraction collector are in use.

Figure 8 Installation of Thermostat and Fraction Collector

1 2



12 Install the tray you have ordered for your fraction collector. The test tube trays can be adjusted in height, depending on the height of the used test tubes. To adjust the height of the trays, press against the snappers at the

Figure 9 Removing the Tray Compartment Divider


1 Installing the Fraction CollectorInstalling a Thermostatted Fraction Collector

guides in the 4 corners of a tray and move the top plate of the tray up or down (see Figure 6 on page 27).

13 Ensure the power switch on the front of the fraction collector is 0FF and the power cables are disconnected.

14 Connect the cable between the fraction collector and the thermostat, see Figure 10 on page 33.

15 Connect the power cables to the power connectors.

16 Connect the CAN cable to other Agilent 1200 Series LC modules.

17 If an Agilent ChemStation is the controller, it must be connect to the LAN interface (should be installed to the detector).

18 Connect the APG remote cable (optional) for non Agilent 1200 Series instruments.

19 Turn ON power by pushing the button at the lower left hand side of the fraction collector.

20 The exhaust fan will turn ON and remove potential solvent vapor from the inside of the instrument. After 2 minutes close the front door. Then the fraction collector will start the hardware initialization process. At the end of this process the status LED should be green.

21 The fraction collector is turned ON when the line power switch is pressed and the green indicator lamp is illuminated. The detector is turned 0FF when the line power switch is protruding and the green light is 0FF.

WARNING Do not disconnect or reconnect the fraction collector to thermostat cable when the power cords are connected to either of the two modules. This will damage the electronics of the modules.

WARNING To disconnect the fraction collector from the line, unplug the power cord. The power supply still uses some power, even if the power switch at the front panel is turned 0FF.

WARNING To avoid personal injury, keep fingers away from the needle area during fraction collector operation. Do not attempt to insert or remove a vial or a plate when the needle is positioned.



Figure 10 Connections at the Rear of the Thermostatted Fraction Collector

Fraction Collector Thermostat cable

CAN-bus AC Power


1 Installing the Fraction CollectorFlow Connections to the Fraction Collector

Flow Connections to the Fraction Collector

Preparation Fraction Collector is installed in the LC system

Parts required Parts from the accessory kit, see “Fraction Collector Accessory Kit Contents G1364-68725” on page 18

WARNING When opening capillary or tube fittings, solvents may leak out. Please observe appropriate safety procedures (for example, goggles, safety gloves and protective clothing) as described in the material handling and safety data sheet supplied by the solvent vendor, especially when toxic or hazardous solvents are used.

WARNING Regularly inspect the inlet / waste tubing assembly and the valve to needle tubing and exchange them if they are worn out or show visible signs of damage.

WARNING Thoroughly follow the described installation procedures to maximize the lifetime of inlet / waste tubing assembly the valve to needle tubing and to avoid potential spills or fraction losses. Regularly inspect the tubings and exchange them if they are worn out or show visible signs of damage.

WARNING This instrument should only be used with solvents that have an ignition temperature higher than 200oC!


Installing the Fraction Collector 1Flow Connections to the Fraction Collector

Route the waste outlet around the instrument as shown below and into a corrugated waste tubing. Fix the waste outlet tubing and the corrugated waste tubing to the instrument with the sticking holders (part of the G1364-68725 accessory kit) as shown below. Route the corrugated waste tubing into a waste container.

Figure 11 Flow Connections to and from the Fraction Collector.

From flow cellTo waste


1 Installing the Fraction CollectorFlow Connections to the Fraction Collector

Figure 12 Fixing and Routing the Waste Tubings


Installing the Fraction Collector 1Fraction Collector Trays

Fraction Collector Trays

Installing the Fraction Collector Trays

1 Press the front door latch-holding button located at the front of the right-side cover.

2 Lift the front door.

3 Adjust the top plate of the test tube tray for the correct tube height if required.

4 Load the fraction collector tray with fraction collector well-plates, test tubes or vials as required.

5 Slide the fraction collector tray into the fraction collector so that the rear of the tray is seated firmly against the rear of the tray area.

6 Press the front of the fraction collector tray down to secure the tray in the fraction collector.

7 Close the front door.

NOTE Installed trays are automatically detected and identified.

NOTE If the tray pops out of position the air channel adapter is not correctly inserted.

NOTE Before starting a run, the instrument has to be correctly configured in the user interface.


1 Installing the Fraction CollectorFraction Collector Trays

Numbering of Vial, Test Tube and Well-plate Positions

With the 4 plates full tray

Plate in the left front position: P1

Plate in the left back position: P2

Plate in the right front position: P3

Plate in the right back position: P4

Vessel: A1; A2;... B1; B2;

With the 2 plates / 10 x 2ml vials or 10 funnels std. trays

Plate in the front position: P1

Plate in the back position: P2

Vessel: A1; A2;... B1; B2;...

Vials / funnels: 1 - 10

With the 100 vials std. tray

Vial: 1 - 100

With the half-trays

Left-hand 40-position tray: 1 - 40

Center 40-position tray: 101-140

Right-hand 40-position tray: 201 - 240

or

Left-hand 15-position tray: 1 - 15

Center 15-position tray: 101-115

Right-hand 15-position tray: 201 - 215


Installing the Fraction Collector 1Fraction Collector Trays

With the 40, 60, 125 or 215 position test tube full trays

Numbering starts in front left corner in columns to the back and then to the right.

WARNING If you are using flammable solvents, remove the plates from the fraction collector after turning it 0FF. You avoid the risk of building explosive gas mixtures in the instrument.

WARNING If you are using flammable solvents, cover the plates. You avoid the risk of building explosive gas mixtures in the instrument.

WARNING Closing mats with adhesive can give some contamination in the system. The adhesive is soluble in most of the solvents used in HPLC.

WARNING In general do not use closing mats with adhesive. The fraction collector has no prepunch needle, therefore the adhesive will clog the needle after several injections.


1 Installing the Fraction CollectorConfigure Well-plate Types

Configure Well-plate Types

If the plate you are using is not found on the “List of Recommended Plates and Closing Mats” on page 168 you may configure a custom plate. Measure the exact dimensions of the plate as marked below and enter the values in the plate configuration table of the ChemStation or the Control Module.

Figure 13 Well-plate Dimensions (straight)

Figure 14 Well-plate Dimensions (staggered)

F

B

C

D

E A

G I

J

column

row

F

B

C

D

E A

G I

J

H


Installing the Fraction Collector 1Configure Well-plate Types

Table 7 Configuring Well-plate Types

Location Description Definition Limits

Rows Number of rows on the plate up to 16

Columns Number of columns on the plate up to 24

Volume Volume (in µI) of a sample vessel

A Row distance Distance (in mm) between the center of two rows

B Column distance Distance (in mm) between the center of two columns

C Plate length X size (in mm) at the bottom of the plate

127.75+/- 0.25 mm (SBS Standard)

D Plate width Y size (in mm) at the bottom of the plate

85.50+/-0.25 mm (SBS Standard)

E Plate height Size (in mm) from the bottom to the top of the plate. If you use well plates with inlets, caps and septa, you have to specify the distance from the bottom to the top of the caps.

up to 47 mm

F Column offset Distance (in mm) from the left edge (bottom) to the center of the first hole (A1)

G Row offset Distance (in mm) from the back edge (bottom) to the center of the first hole (A1)

H Column shift Offset (in mm) to Y when the rows are not straight but staggered

J WeIl depth Distance (in mm) from the top of the plate to the bottom of the well. If you use well plates with inlets, caps and septa, you have to specify the distance from the top of the septa to the bottom of the inlets.

up to 45 mm


1 Installing the Fraction CollectorConfigure Well-plate Types

Well X size Size of the well in x direction (Plate length) If you use well plates with inlets, caps and septa, you have to specify the x size of the septa.

min. 3.7 mmmin. 3.0 mm with position accuracy alignment (micro scale)

Well Y size Size of the well in y direction (Plate width). If you use well plates with inlets, caps and septa, you have to specify the y size of the septa.

down to 3.7 mmmin. 3.0 mm with position accuracy alignment (micro scale)

Bottom size For round wells, the relative of the top and bottom of the well

1.0: cylindrical well0.0: conical well

Square Click in the field to specify whether the well is rectangular or round

Yes: rectangularNo: round /oval

Is well plate Click in this field to specify if this is a well plate or not. Relevant for continuous flow operation.

Yes: well plate or MALDI TargetNo: Vial Tray or Eppendorf tray

Table 7 Configuring Well-plate Types (continued)

Location Description Definition Limits

NOTE The distances need to be measured with high precision. It is recommended to use a caliper.

If possible contact the vendor of non-predefined well plates to obtain these dimensions.


Installing the Fraction Collector 1Transporting the Fraction Collector

Transporting the Fraction Collector

When moving the fraction collector inside the laboratory, no special precautions are needed. However, if the fraction collector needs to be shipped to another location via carrier, ensure:

✔ The transport assembly is in the park position. Use the ChemStation or the Control Module for this command.

To move the arm to the park position:

1 Switch to the Diagnosis view of the ChemStation and select Fraction Collector > Maintenance Positions. from the Maintenance menu

2 In the upcoming dialog box click Park Arm.

✔ The vial tray and the sample transport mechanism is secured with the transport protection foam.


1 Installing the Fraction CollectorTransporting the Fraction Collector



2Configuration and Operation of the Fraction Collector

Configuration of the Fraction Collector 46

Delay volumes and delay calibration 52

Setting up a Fraction Collector Method 67

Starting your run with fraction collection 74

Viewing your Results 78

Special Applications 80

Limitations and how to avoid problems 85

Check-out Procedures 86

Application Notes 91


2 Configuration and Operation of the Fraction CollectorConfiguration of the Fraction Collector

Configuration of the Fraction Collector

Configuration of the fraction collector in the ChemStation

In order to setup or change the configuration parameters of your fraction collector select More Fraction Collector > Configuration from the Instrument menu or right-click on the fraction collector icon in the graphical user interface. In the resulting Fraction Collector Configuration dialog box (Figure 15 on page 47) the configuration of the Trays, the Fraction Delay Volumes, the Collection Order, the Needle Movement and the Well-Plates can be specified.

Trays

In the online mode of the Agilent ChemStation the type of the installed tray is recognized automatically. In the off-line mode an appropriate tray type can be chosen from a dropdown list. For a selection of supported trays see “Supported Trays for Fraction Collectors” on page 162.

Tube volume [ml] Volume of the test tube should be specified 10% less than the maximum fill volume to avoid overfill.

Tube height [mm] The exact test tube height has to be specified. For preparative scale fraction collector the minimum height is 48 mm and the maximum height is 100 mm. For the analytical scale fraction collector with 50 mm needle (standard) the maximum height is 48 mm. By changing to the short 20 mm needle the maximum tube height can be in creased to 75 mm. For the part numbers of the different needle assemblies see “Needle Assemblies” on page 173.

CAUTION Before using the preparative scale fraction collector G1364B the delay calibration adapter G1364-87301 has to be removed from the needle carrier assembly.

NOTE In the online ChemStation the installed tray is recognized and the default settings for tube volume and tube height are loaded and displayed.


Configuration and Operation of the Fraction Collector 2Configuration of the Fraction Collector

Use for Sample Recovery This checkbox only appears, if multiple fraction collectors are configured (see Figure 34 on page 81). Then the last fraction collector can be selected for sample recovery. If four fraction collectors are configured, the fourth one will automatically used for sample recovery. To change the order of your fraction collectors select Configure 1200 Access from the Instrument menu. In the Configuration dialog box use the Up or Down button to change the configuration order and restart your ChemStation.

Figure 15 Fraction Collector Configuration dialog



Fraction Delay Volumes

The Fraction Delay Volume specifies the volume between the detector cell and fraction collector diverter valve. This volume has to be determined during the delay calibration. See “Delay volumes and delay calibration” on page 52 for a detailed description.

Collection Order

The Collection Order describes the way of capillary movement during fraction collection. Four different settings are possible:

• row-by-row

• column-by-column

• shortest way by row

• shortest way by column

Shortest way by row/column is recommend, if Continuous flow is selected as Collection mode.

Collection Mode

The Collection Mode defines the triggering of the diverter valve. For Discrete fractions the diverter valve switches to waste while the needle moves to the next test tube, vial or well. For Continuous flow the diverter valve is not switching during needle movements, except for long movements in the row-by-row or column-by-column mode. In addition the needle cannot move Into location during continuous flow operation. Continuous flow is only available for well plates.

Needle Movement

Into location In the Into Location mode the needle tip moves into the well to the specified Depth (in mm). This option is only available for the analytical scale fraction collector in the Discrete fractions mode.

Above location In the Above Location mode the needle tip stays at the specified Distance (in mm) above the well during fraction collection.

Contact Control In this mode the needle tip moves down to the well bottom until is reaches the specified Distance (in mm) between needle tip and the vial/well bottom. This ensures that the forming droplet is in contact to the



vial/well bottom. During the continuing filling process the needle tip moves upwards while staying in contact with the surface of the collected liquid. This option is recommend for low flow rates and small fraction volumes to avoid air bubbles and accomplish an accurate fraction volume. It is only available for the analytical scale fraction collector.

Well-Plates

In the Well-Plates section the type of well plates used in a well plate tray can be configured. The well plate type can be chosen from the Plate Type dropdown list. More detailed information about all pre-configured well plates can be found in the Instrument menu. Only one type of well plate can be used on the well plate tray.

The checkbox Treat as 4 x 96 is available only for 384 well plates and allows to split the 384 well plate virtually into 4 separate 96-well plates. This only changes the collection order. The numbering of the wells remains as indicated on the plate. The four virtual 96 well plates start at locations A1, A13, I1 and I13, respectively.

The filling order of each quarter is as specified in the Collection Order section. When the 384 well plate is split into four equal quarters the order of the four plates is the same as displayed in the Installed Plates section.

In the Reserved Locations, you have the possibility to specify locations that will not be used for Fraction Collection (see Table 8).

Table 8 Syntax for the definition of Reserved Locations

Location Syntax Description

Rows A Row A can’t be used

A,B Rows A and B can’t be used

A-D A, B, C and D can’t be used

A-D,F Rows A, B, C, D and F can’t be used

Columns 1 Column 1 can’t be used

1,2 Columns 1, and2 can’t be used

1-4 Columns 1, 2, 3 and 4 can’t be used



Configuring multiple fraction collectors

To increase the capacity of the systems up to three fraction collectors can be configured by using the Agilent 1200 Series 12-position13-port valve G1160A. In addition a fourth fraction collector can be configured for recovery collection.

• The Configuration has to be edited for all fraction collectors. The last fraction collector in the configuration can be selected for sample recovery. To configure the recovery fraction collector please read “Sample Recovery” on page 81.

• With multiple fraction collectors a fraction start location has to be specified to define the starting position. For further details see “Assignment of Start Location for fraction collection” on page 74. Never used the valve settings to define a start location on systems with multiple fraction collectors.

• We recommend to use inlet tubing of the same length for all fraction collectors. Otherwise a Delay Calibration has to be performed for each of those fraction collectors. For the recovery fraction collector the delay volume parameter will be ignored.

1-4,12 Columns 1, 2, 3, 4 and 12 can’t be used

Single locations G12,H12 Locations G12 and H12 can’t be used

Table 8 Syntax for the definition of Reserved Locations (continued)

Location Syntax Description

NOTE The Delay Calibration can only be performed for fraction collector 1 of your configured system.

The calculated delay volume is used for all fraction collectors; therefore, the volume (tubing) between the selection valve and each installed fraction collector must be identical.

Otherwise fraction collector 2 and fraction collector 3 have to be temporarily configured as fraction collector 1 during the delay calibration.



• The fraction collector inlet tubings have to be connected to port 1-3 of the selection valve in the same order as configured in the ChemStation. The recovery fraction collector is always connected to the waste tubing of the main fraction collectors. The waste tubing from multiple fraction collectors and the inlet tubing to the recovery collector have to be connected through a T-Piece.

• Fraction Collector 4 will always be used as a recovery fraction collector.


2 Configuration and Operation of the Fraction CollectorDelay volumes and delay calibration

Delay volumes and delay calibration

Delay times and volumes

Figure 16 shows a schematic drawing of the flow path between the detector and the fraction collector with the two delay volumes VD1 and VD2. For peak-based fraction collection the system delay times tD1 and tD2 can be calculated by dividing the delay volumes by the flow rate .

The delay volume VD2 is a system parameter, it depends on the installed fraction collector tubing and the fraction collector needle. It is approximately 23 µl for the fraction collector AS and approximately 120 µl for the fraction collector PS. Delay volume VD1, which is specified in the Fraction Collector Configuration window, is determined using the Delay Volume Calibration feature of the ChemStation software.

When a peak is detected during a purification run (Figure 17) the diverter valve is triggered using the following delay time calculations:

Start of fraction collection: t = t0 + tD1

End of fraction collection: t = tE + tD1 + tD2

Figure 16 Delay Volumes and delay times

ν·


Configuration and Operation of the Fraction Collector 2Delay volumes and delay calibration

Detector signal delay

Every Agilent 1200 Series detector that is used for triggering fractions has an internal signal delay caused by filtering the raw data. The signal delay depends on the Peakwidth setting of the detector and is accounted for when the diverter valve is triggered. Table 9 and Table 11 list the internal signal delay times for different Peakwidth settings.

Figure 17 Chromatogram with peak start t0 and peak end tE

Table 9 Signal delay times for the Agilent 1200 Series DAD/MWD (G1315B/G1365B)

Peakwidth (min) Response time (sec) Signal delay (sec)

<0.01 0.1 0.05

>0.01 0.2 0.15

>0.03 0.5 0.5

>0.05 1.0 1.25

>0.10 2.0 2.75

>0.20 4.0 5.9

>0.40 8.0 11.9

>0.85 16.0 23.9



Table 10 Signal delay times for the Agilent 1200 Series DAD/MVD SL (1315C/G1365C)


<0.0025 0.02 0.0375

>0.0025 0.05 0.0625

>0.005 0.1 0.144

>0.01 0.2 0.294

>0.03 0.5 0.619

>0.05 1.0 1.27

>0.1 2.0 2.57

>0.2 4.0 5.17

>0.4 8.0 10.4

>0.85 16.0 20.8

Table 11 Signal delay times for the Agilent 1200 Series VWD G1314B


<0.005 <0.12 0.07

>0.005 0.12 0.14

>0.01 0.25 0.29

>0.025 0.5 0.58

0.05 1 1.3

0.1 2 2.8

0.2 4 6.0

0.4 8 12.3



Table 12 Signal delay times for the Agilent 1200 Series VWD SL G1314C


<0.00125 <0.02 0.0182

>0.00125 0.02 0.0364

>0.0025 0.05 0.0728

>0.005 0.12 0.146

>0.01 0.25 0.328

>0.025 0.5 0.710

>0.05 1 1.49

>0.1 2 3.08

>0.2 4 6.26

>0.4 8 12.6

Table 13 Signal delay times for the Agilent 1200 Series FLD G1321A


<0.005 <0.12 0.018

>0.005 0.12 0.073

>0.01 0.25 0.18

>0.03 0.5 0.44

>0.05 1 0.96

>0.1 2 2.0

>0.2 4 4.2

>0.4 8 8.6



The stop-time of the run in the ChemStation must be set to at least:

Total duration of time table (time of last entry OFF) + fraction collector delay time for time-based fraction collection.

End of last peak (tE) + fraction collector delay time for peak-based fraction collection

CAUTION If the internal signal delay is longer than the delay time tD1 some of the peak will be lost. The maximum allowed signal delay time can be calculated using the following equation:

After calculating the maximum signal delay time a Peakwidth setting can be selected that gives a signal delay time, which is shorter than the calculated maximum signal delay time. This Peakwidth setting should then be used for the LC purification run.

Signal delay time max( )VD1

ν·----------= ν· Flow rate=

NOTE We recommend to set the Peakwidth always to > 0.01 for the DAD and MWD or to > 0.005 for the VWD.

If the Peakwidth setting cannot be reduced and the signal delay time is longer than tD1 it is also possible to increase VD1 by adding additional tubing. However this higher delay volume will increase the peak dispersion between detector and fraction collector.

VD1 ν·⁄( ) 0.1min+

VD1 ν·⁄( ) 0.1min+



Performing a delay calibration with an UV detector

1 Place a vial containing the Delay Sensor Calibrant (Part No. G1946-85020) in position 1 of the Autosampler.

2 Remove the installed column and connect the capillaries with a zero-dead-volume connector.

3 Install the 40 x 2ml vial tray (analytical scale) or the 15 x 6ml vial tray (preparative scale) in the fraction collector.

4 Connect a bottle of water to Channel A.

5 Switch to Diagnosis View (if necessary).

6 Open the AFC Delay Volume Calibration status window (see Figure 18) from the Fraction Collector sub-menu of the Maintenance menu:

Maintenance > Fraction Collector > Delay Volume Calibration

7 The Info field gives the instruction to turn ON the pumps (see Figure 18). Click Continue.

Figure 18 Delay Volume Calibration status window



When you have turned ON the pumps, click Continue again to start the delay volume calibration.

The method to be used for the delay volume calibration is displayed (DGCALAS.M is the default calibration method for the analytical scale fraction collector and DGCALPS.M is the default method for the preparative fraction collector); if you want to change to a different method, click the Change Method button and select a new method. See also “Making Your Own Calibration Method” on page 66

The Info field shows the current status of the delay volume calibration, and gives instructions and warnings.

8 Install the delay calibration adapter G1364-87301 by attaching it to the needle carrier assembly (G1364B preparative scale only).

The selected method is loaded, and the delay volume calibration sample is injected.

9 After the delay calibration has been finished, you are prompted to remove the delay calibration adapter (G1364B preparative scale only).

10 When the calibration run has finished, click OK. The Calibration Results panel (see Figure 20) is displayed.

Figure 19 AFC Delay Volume Calibration Window



The Calibration Results panel contains a results table and the acquired chromatograms from the detector(s) and the fraction collector delay sensor. The results table contains

• the name and serial number of the detector,

• the current delay volume (stored in the fraction collector configuration),

• the calculated delay volume from the calibration.

To print the Calibration Results panel click Print.

11 If you want to store the calculated delay volume in the fraction collector configuration, click Calibrate. A confirmation dialog box is displayed when the delay volume has been stored; to close the confirmation click OK.

If you do not want to store the calculated delay volume in the fraction collector configuration, click the Close button. A confirmation dialog box is displayed; click OK to close, the Confirmation or Cancel to return to the Calibration Results panel.

Figure 20 Delay Calibration Results Panel



NOTE The Delay Calibration can only be performed for fraction collector 1 of your configured system.

The calculated delay volume is used for all fraction collectors; therefore, the volume (tubing) between the selection valve and each installed fraction collector must be identical.

Otherwise fraction collector 2 and fraction collector 3 have to be temporarily configured as fraction collector 1 during the delay calibration.




Performing a delay calibration with an MSD

1 Place a vial with the Delay Sensor calibrant (Part No. G1946-85020) in position 1 of the Autosampler.

2 Remove the installed column and connect the capillaries with a zero-dead-volume connector or a mixer (Part No. 79835-87330).

3 Install the 40 x 2ml vial tray (analytical scale) or the 15 x 6ml vial tray (preparative scale) in the fraction collector.

4 Connect a bottle of water to Channel A of the main pump.

5 Connect a bottle of water with 0.1% acetic acid or ammonium formate to the make-up pump.

6 Load the method DGCALAS.M (analytical scale) or DGCALPS.M (preparative scale) and adjust the flow in the main pump to the flow you will be using for the analysis.

7 Set the flow of the make-up pump to the flow rate you will be using for your analysis.

8 Set the active splitter to the split ratio as used during operation.

9 Save the method to a new name.

10 Go to Diagnostics menu; select Delay volume Calibration from the Fraction Collector Sub-menu within the Maintenance menu.

11 Press Change method... to select the method you created. The new method name appears on the info line.

12 Start the active splitter.

13 Press Continue to execute the Delay Volume Calibration

14 Install the Delay Calibration Adapter G1364-87301 by attaching it to the needle carrier assembly (G1364B preparative scale only).

CAUTION Unlike for UV detectors, the delay calibration for the MSD needs to be performed whenever the flow rate is changed.



The selected method is loaded and the Delay Calibration Sample Injected.

15 After the delay calibration has been finished, you are prompted to remove the delay calibration adapter (G1364B preparative scale only).

16 When the Calibration Run is finished, press OK.

17 Examine the value for the MSD Calculated Delay time.



The delay time should be at least 5 seconds but may, in fact, be negative (see Figure 21).

18 If the measured delay time is greater than 5 seconds, you can skip to step 23, otherwise, continue with step 19.

19 Determine how much additional time is needed to create a 5-second delay. For example, if the time displayed is -1 sec, a six-second delay is needed.

20 Using the main pump flow rate, determine the desired additional delay volume. For example, at 3 ml/min and a six-second delay, an additional 3.0 x 6/60 or 0.3 ml is required.

21 Cut off a sufficient piece of the supplied ETFE tubing (Part No. G1968-60500) to provide the needed volume. The tubing's volume is 8.1 µl/cm. In this case, 37 cm would be required.

22 Add the tubing between the splitter and the fraction collector or, if multiple fraction collectors are installed, between the splitter and the G1160A valve.

23 Perform the calibration again. Verify the time is sufficient (>5 seconds, see Figure 22).

Figure 21 Delay Volume Calibration results

CAUTION If the measured delay time is significantly greater than 5 seconds, you should continue with the calibration at step 16 to avoid losing fractions at the end of the run.



24 Press Calibrate if you want to store the new calculated delay volume for the UV detector in the Fraction Collector Configuration (see also “Configuration of the fraction collector in the ChemStation” on page 46).

25 Press Print to get a hard copy of the report.

26 Stop the splitter.

27 Return to the Method and Run Control View and load the method you will be using to run your samples. Select Fraction Collection from the More MSD... sub-menu under the Instrument menu

Press Parameters and enter the final calculated delay time that appears in the report in the Collector Delay field, first converting it to minutes. In this case you would enter 11.991/60 = 0.20 minutes (see Figure 23).

Figure 22 Delay time greater than 5 seconds



28 Save the method.

Figure 23 Fraction Collection Parameters dialog box with delay volume entry

NOTE If you anticipate running samples at a variety of flow rates, it is best to calibrate at the highest likely flow rate first and install the appropriate length tubing. This way, there will be sufficient delay for the lower flow rates and new lengths will not need to be cut.



Making Your Own Calibration Method

You make your own calibration method by editing one of the default calibration methods:

1 Load the default calibration method: DGCALAS.M in the case of an analytical system and DGCALPS.M in the case of a preparative system.

2 Change the method parameters (flow, runtime, solvent composition, injection volume or detector parameters) to suit your adapted calibration procedure.

3 Save the method with a new name in the method folder for your instrument.

4 Follow the appropriate procedure as described in the previous sections for running the method.



Configuration and Operation of the Fraction Collector 2Setting up a Fraction Collector Method

Setting up a Fraction Collector Method

In order to setup or change the method parameters of your fraction collector select Setup Fraction Collector from the Instrument menu or right-click on the fraction collector icon in the graphical user interface. This will open the Setup Fraction Collector dialog box as displayed in Figure 24. In the Setup Fraction Collector dialog box general method settings are specified.

Figure 24 Set up Fraction Collector dialog box


2 Configuration and Operation of the Fraction CollectorSetting up a Fraction Collector Method

Fraction Trigger Mode

Use Timetable Enables the Timetable

Peak-based If Peak-based is selected, the collection of a fraction is triggered by the signal of the detector, e.g. the Agilent 1200 Series diode array detector or variable wavelength detector. The detailed trigger conditions are specified in the Peak Detectors table. In the peak-based trigger mode all entries in the Timetable are ignored.

Max. Peak Duration Defines a maximum collection time in case that the signal does not reach the condition to cut the fraction as exhibited in Figure 25. The could be caused by tailing peaks or if the baseline is drifting during gradient runs. The default value is set to 0.5 minutes. If broad peaks are expected, this value should be increased without exceeding the run time.

Peak Detectors

In the Peak Detectors section a list of all Peak Detectors that are connected to the system is displayed. Agilent 1200 Series diode-array detectors, multi- wavelength detectors, variable wavelength detectors and fluorescence

Figure 25 Maximum Fraction Duration

Threshold

Max. Peak Duration



detectors are recognized automatically. Other Detectors, e.g. Agilent 6000 Series mass-selective detectors or HP1050 Detectors, are connected through the Universal Interface Box (UIB).

The Peak detector table contains seven columns:

Working Mode For each peak detector Threshold only, Threshold/Slope or Slope only are possible.

In the Threshold only mode the settings for Up Slope, Down Slope and Upper Threshold in the subsequent columns are ignored. Fraction collection is triggered whenever the detector signal exceeds the specified threshold value. When the signal drops below the threshold value fraction collection is stopped.

In the Slope only mode fraction collection is triggered on the slope of the detector signal. Adequate values for Up Slope and Down Slope can be specified in the corresponding fields.

In the Threshold/Slope mode fraction collection is triggered on the corresponding values for threshold and slope. The fraction collection is started if the detector signal exceeds both the threshold and the Up Slope value. The fraction collection is stopped if the detector signal drops either below the threshold or the Down Slope value.

To specify the trigger values Up Slope, Down Slope, Threshold and Upper Threshold we recommend to use the Fraction Preview tool as described in “Fraction Preview” on page 72.

Upper Threshold At high absorbance values the light intensity on the detector is extremely low and consequently detector noise will be superimposed on the detector signal. In this case the detector noise might trigger fraction collection. To avoid false fraction collection triggering, we recommend setting an Upper Threshold well below the limit where this false triggering effect might occur. As soon as the detector signal exceeds the Upper Threshold, settings for Up Slope or Down Slope will be ignored until the signal drops again below the Upper Threshold.

When using more than one peak detector fraction collection can be triggered either when all selected peak detectors detect a peak or when at least one selected peak detector detects a peak basing on the settings in the Peak Detectors table above.



If an MSD is used for mass-based fraction collection, Use MSD for mass-based Fraction Collection must be checked.

Timetable

The Timetable can be used to program changes in the Fraction Trigger Mode during the analysis by entering a Time and specifying the trigger settings.

Trigger Mode Off, Peak Based and Time Based can be selected. If the Off is selected, no fractions are collected. The last entry in the timetable has to be the command Off.

Whenever the Peak Based mode is specified fractions will be collected based on the peak detection parameters given in the Peak Detector table. Additionally a Maximum Peak Duration in minutes has to be specified. This parameter is mandatory if you use Peak Controlled fraction collection, but is disabled for Time Based fraction collection.

When the Time Based mode is chosen two different options are available:

• The # of Fractions can be edited to collect a fixed number of equal fractions in a give time interval. This time interval is defined by the time value in the current and following timetable line.

• Timeslices [min] can be edited to collect fractions with a defined collection time. With this option the collection time of the last fraction can be shorter. This depends on the overall runtime.

For editing the Timetable the functions Insert, Append, Cut, Copy and Paste are offered.

To access the additional sections in the Setup Fraction Collector dialog box click More.

Time

In the time section of the dialog box the Stoptime and the Posttime for the fraction collector can be specified. By default the Stoptime is set to as pump and the posttime is switched OFF.

Rinse Fraction Collection Needle (Analytical Scale only)

If Discrete Fractions is selected as Collection Mode (see also“Collection Mode” on page 48), you can setup a needle rinse step before the fraction collection and/or between fractions. Then the needle will move to the funnel



on the internal tray and the diverter valve will switch to flush the needle in order to avoid carry over from the previous fraction. The instrument will determine, if it is possible to rinse the needle before the next fraction is expected.

If you have recovery positions in your fraction collector or if you are using one fraction collector for sample recovery in a multiple fraction collector configuration, the function rinse between fractions is ignored.

Auxiliary

In the Auxiliary section the Maximum fill volume per location can be specified. If as configured is selected, the pre-configured volume (see Instrument > Pre-configured Wellplate Types) is used. This ensures that the location (well, vial or tube) cannot be overfilled during fraction collection. This volume can be further reduced by defining a customized volume.

Additional check boxes in this section provide the opportunity to Store the temperature signal and the UIB signal.



Fraction Preview

To determine the appropriate fraction collection parameters the Agilent ChemStation provides a valuable tool that becomes accessible by pushing the button labelled Fraction Preview Tool (Figure 26) in the Peak Detectors section.

The Fraction Preview screen allows to test the fraction collection parameters against an example chromatogram. It can also be used to optimize the fraction collection parameters interactively. With the help of this tool values for up and down slope as well as for upper and lower threshold can easily be graphically specified. To load a chromatogram e.g. a pilot run click Load Signal. Parameters can now be changed either manually in the Detector Table and Timetable or graphically in the Fraction Preview screen. By pushing the desired buttons on the right hand side of Fraction Preview screen the chromatogram can be

Figure 26 Fraction Preview dialog box



zoomed, the values for up and down slope can be specified and the upper and lower threshold level can be set-up. The graphically specified values are automatically transferred to the Peak Detector Table.


2 Configuration and Operation of the Fraction CollectorStarting your run with fraction collection

Starting your run with fraction collection

Resetting the current fill levels

To protect the collected fractions against contaminations and to avoid overfilling of single positions the Agilent 1200 Series fraction collector stores current fill levels. It will block used positions, if no fraction start location has been specified.If a new tray of the same type is inserted in the fraction collector, all fill levels can be reset (not just the well plate, vial or test tube). After re-installation of the tray a pop-up window as displayed in Figure 27 will be displayed. Click on Yes to reset all fill levels from the previous run.

Assignment of Start Location for fraction collection

The start location for fraction collection can either be assigned in the Sample Info (Figure 28) dialog box in the RunControl menu, in the Sequence Parameters (Figure 29) dialog box or in the Sequence Table in the Sequence menu. All fraction start locations are entered in the following formats.

Figure 27 Reset fill volumes

CAUTION The window (Figure 27) to reset current fill levels is not displayed, if the well plate, the vial or the test tube is removed without removing the complete tray. Consequently contaminated fractions, unwanted overfill events or split fractions might be observed during the following LC run.


Configuration and Operation of the Fraction Collector 2Starting your run with fraction collection

For vials and test tubes:

fraction collector # - vessel number, e.g. 1-Vial 1

For well plates:

fraction collector # - plate number - row - column, e.g. 1-P1-A-01

withfraction collector # = 1,2; plate number = P1-P4; row is A to H and column = 1-12 for 96-well plates.

All settings made in the Sequence Table overrule the settings made in the Sequence Parameters screen. In addition to the exact position for fraction collection start in the Sequence Table it is also possible to specify Next Plate and Next Location. In the former case fraction collection is started at the next free plate and in the latter case fraction collection is started at the next free location.

If a fraction start location is specified, the collector will start at this location, even if this vessel has been used during a previous run.

NOTE Note the orientation of the well plate. The starting position A1 is always in the back left corner of the plate.

Figure 28 Sample Info dialog box


2 Configuration and Operation of the Fraction CollectorStarting your run with fraction collection

If no fraction start location has been specified, the fraction collection starts at the first empty vessel or location.

To start a single run

• click on the Start button in the graphical user interface of the ChemStation.

• select Run Method from the RunControl menu.

• press F5

To start the sequence

• click on the Start button in the graphical user interface of the ChemStation.

• select Run Sequence from the RunControl menu.

• press F6

CAUTION With multiple fraction collectors a fraction start location must be specified, e.g. 1-P1-A-01 or 2-Vial 10. Otherwise the fraction collection of the next run will start at the fraction collector, where the last run stopped. Never used the valve settings to define a start location.

Figure 29 Start Location in Sequence Parameters dialog box


Configuration and Operation of the Fraction Collector 2Starting your run with fraction collection

Online tick marks

To display tick marks for fraction start and stop events in you Online Plot click on the Change button in your Online Plot window. Then check Show fraction collection ticks in the Edit Signal Plot window (Figure 30).

Figure 30 Online Tick Marks


2 Configuration and Operation of the Fraction CollectorViewing your Results

Viewing your Results

Data Analysis

In order to display the tick marks for the collected fractions on the screen, click Signal options from the Graphics menu. Then choose Separated in the Layout dropdown list.

To review your chromatograms, file information and a fraction list, select the Data Analysis view and press Fraction Task as displayed in Figure 31.

Figure 31 Fraction Task button

Chromatogram with Fraction Tick Purification Task

Marks

Fraction table


Configuration and Operation of the Fraction Collector 2Viewing your Results

In order to display the tick marks for the collected fractions on the screen, click Signal options from the Graphics menu. Then choose Separated in the Layout dropdown list.

Report

In order to create reports with a fraction table and tick marks the Specify Report box the item Add Fraction Table and Tick Marks has to be checked.

Figure 32 Fraction collection report setup


2 Configuration and Operation of the Fraction CollectorSpecial Applications

Special Applications

Pooling

We define Pooling as collecting fractions from multiple injections into the same fraction locations. In the Agilent Chemstation, there are two ways to initiate pooling:

• If you want to collect multiple injections from the one sample vial or sample well into the same fraction location, specify multiple injections in the Sequence Table.

• In case you want to collect multiple injections from the several sample vials or sample wells into the same fraction location, specify the same Fract. Start for multiple samples in the Sequence Table. An example is displayed in Figure 33 on page 80. In this sequence the Sample 1 will be injected from Plate 1 Position A1 of the Agilent 1200 Series high performance autosampler and fraction collection will start at Plate 1 Position A1 of the fraction collector.For the next Sample 2, which is injected from a different location in the high performance autosampler, the fraction collection will start again at Plate 1 Position A1 of the fraction collector.

Figure 33 Pooling Sequence

CAUTION If you specify multiple injections for pooling, the overfill protection is not activated. It is the user's responsibility to make sure that all fraction collection locations are large enough to completely collect all pooled fractions. If a fraction collection location is overfilled, an error message occurs and the pump is shut OFF.


Configuration and Operation of the Fraction Collector 2Special Applications

Sample Recovery

The Agilent 1200 Series fraction collectors offers different possibilities for sample recovery:

• The preferred recovery strategy is to install multiple fraction collectors in your LC systems and use the last of those fraction collectors for recovery. This recovery fraction collector can be selected in the Configuration dialog box as displayed in Figure 34. The fourth fraction collector in your systems will always be used for sample recovery. Also read “Configuring multiple fraction collectors” on page 50.

• For the following tray configurations fixed recovery location will be assigned from the ChemStation. In order to disable the recovery the tray configuration has to be changed.

1 Standard tray for two well plates + 10 2ml x vials (G2258-60011) and half tray for 40 x 2 ml vials (G1313-44512).In this configuration the forty 2 ml vials on the half will automatically be used for recovery.

2 Standard tray for two well plates + 10 x 2ml vials (G2258-60011) and half tray for 15x 6 ml vials (G1313-44513).In this configuration the fifteen 6 ml vials on the half will automatically be used for recovery.

3 Standard tray for two well plates + 10 x 2ml vials (G2258-60011) and half tray with 40 funnels (G1364-84532).In this configuration the funnels on the half will automatically be used for recovery.

• The Standard tray for two well plates + 10 funnels (G1364-84522) allows two modes of operation.

Figure 34 Sample Recovery Fraction Collector


2 Configuration and Operation of the Fraction CollectorSpecial Applications

1 If well-plates are configured in the fraction collector configuration the (checkbox Plate 1 or Plate 2 are selected), the 10 funnel locations will automatically be used for recovery.

2 If the checkbox Plate 1 and Plate 2 are not selected, the 10 funnel position can be used for regular fraction collection.

Semi-preparative operation

The analytical scale fraction collector G1364C can be modified for semi-preparative operation. This allows to use of external collection vessels through funnels at high flow rates above 10 ml/min. The maximum flow rate depends on the viscosity of the solvent.

In order to modify the analytical fraction collector the following parts have to be installed:

• 20 mm needle assembly (G1364-87202)

• preparative tubing kit (G1364-68711)

• internal tray preparative scale (G1364-63113)

The procedures to exchange these parts are described in“Replacing the Inlet/Waste Tubings” on page 141, “Replacing the Valve to Needle Tubing” on page 145, “Exchanging the Analytical Needle Assembly” on page 150, and “Exchanging the Internal Tray” on page 156.

In addition the wellplate adapter has to be removed for operation with the shorter needle.

CAUTION With each start of a new sequence the recovery will start at the same positions. In order to avoid contamination the vessels that contain the recovery fractions have to be exchanged.

CAUTION The number of recovery locations automatically defines the maximum number of injections. When using the standard tray for two well plates and 10 funnels, only ten injections per sequence are executed.


Configuration and Operation of the Fraction Collector 2Special Applications

In the ChemStation Software the new configuration has to be specified in order to consider the new volumes for the delay calibration. In the Configuration dialog box of the fraction collector select More. Then change the Inner Capillary Diameter to 0.8 mm as well as the Needle Type to semi-prep needle as displayed in Figure 35 on page 83. For further information on delay volumes and delay volume calibration see “Delay volumes and delay calibration” on page 52.

Using high test tubes in the analytical scale fraction collector

In the standard configuration the analytical scale fraction collector G1364C the maximum test tube height is limited to 48 mm. This limitation can be extended to 75 mm if

• the standard 50 mm needle assembly (G1367-87200) is replaced by the20 mm needle assembly (G1364-87202)

• the wellplate adapter is removed.

Figure 35 Fraction collector configuration parameters for semi-prep operation


2 Configuration and Operation of the Fraction CollectorOptimizing fraction collection

Optimizing fraction collection

Time-basedfraction

collection

• Time slices must have a length of at least 0.05 min.

• Set # of Fractions such that length of resulting fractions is at least 0.05 min.

Peak-basedfraction

collection

• Set threshold and slope values such that length of fractions is at least 0.05 min.

• Unresolved peaks can be separated using appropriate threshold and slope values. If two unresolved peaks are to be collected as one fraction, collect based on threshold only.

• If the baseline of the chromatogram is below or above 0 mAU, this offset is not accounted for when triggering peaks using a threshold value. The threshold value is always added to 0 mAU.


Configuration and Operation of the Fraction Collector 2Limitations and how to avoid problems

Limitations and how to avoid problems

Rinse FractionCollection Needle

If Rinse Fraction Collection Needle is set to Between fraction collection, at least 0.3 min are required to perform this task. When doing time-based fraction collection rinsing the needle is only possible between two time table entries, which must have a gap of at least 0.3 min. For peak-based fraction collection a time gap of also at least 0.3 min is required. If a new peak is detected during the rinse process, it is aborted and the needle moves back to the next free fraction position. Depending on flow rate and delay volume VD1 the beginning of this peak may be lost.

If you have recovery positions in your fraction collector or if you are using one fraction collector for sample recovery in a multiple fraction collector configuration, the rinse function between fraction collection is ignored.

NeedleMovement

The option into location under Needle Movement in the fraction collector configuration must only be used for capped 2 or 5 ml vials or well-plates. Using other or open vials with this command can lead to a Movement failed error.

Replacingfraction

containers

When replacing filled tubes, vials or well-plates from the fraction collector make sure to remove and re-insert the complete tray. Otherwise the fraction collector will not recognize that the fraction containers were emptied.

Pooling • When pooling fractions, overfill protection no longer exists. It is the user's responsibility to make sure that all fraction collection locations are large enough to completely collect all pooled fractions. If a fraction collection location is overfilled, an error message occurs and the pump is shut OFF.


2 Configuration and Operation of the Fraction CollectorCheck-out Procedures

Check-out Procedures

The purpose of the check-out injection is the verification of the correct installation and configuration of the Agilent 1200 Series purification system. In addition the checkout procedure can used for troubleshooting, if the Agilent 1200 Series fraction collector doesn’t operate as expected.

The checkout procedure depends on the flow range (analytical scale AS or preparative scale PS) and on the peak trigger (UV-based or mass-based).

In the following sections the check-out procedures for the UV based trigger are described. For the mass-based system refer to the Agilent 1200 Series Purification system manual G2262-90010.

Check-out procedure for UV-triggered fraction collection AS

Prerequisites

✔ The Agilent 1200 Series LC system has been flushed thoroughly with the liquid phase as described in Table 14 and the lamp(s) of the UV detector have been switched on for at least 15 minutes to provide a stable baseline.

✔ The Agilent 1200 Series fraction collector has been installed and configured. The delay volume has been calibrated as described in “Performing a delay calibration with an UV detector” on page 57.

✔ The 40-vial tray (G1313-44512) with at least four 2ml vials is installed.

✔ All wells are empty and the fill levels, which are stored on the module, have been reset. To verify this remove and insert the 40-vial tray. Then click on Yes in displayed dialog box.


Configuration and Operation of the Fraction Collector 2Check-out Procedures

Table 14 Method parameters for UV-triggered fraction collection check-out AS

Agilent 1200 Series modules • standard or micro degasser• isocratic, quaternary or binary pump• autosampler/ well plate sampler• column compartment• diode-array detector, multi-wavelength detector or variable

wavelength detector with analytical flow cell• fraction collector analytical scale

Column ZORBAX Eclipse XDB-C18, 5 µm, 150 x 4.6mm (993967-906) or equivalent

Sample Check-out sample (01080-68704)

Flow 0.8 ml/min

Stoptime 6 minutes

Solvent A 10% (HPLC grade water)

Solvent B 90% (HPLC grade acetonitrile)

Wavelength VWD 254 nm

Wavelength DAD/MWD Signal A: 254/16nm, Reference: 360/80nm

Peakwidth (Responsetime) > 0.1min (2s)

Injection volume 1 µl

Column temperature 40.0 0C

Fraction trigger mode Peak-based

Max. peak duration 0.5 min

DAD/MWD working mode Threshold/slope

Upslope 5 mAU/s

Downslope 10 mAU/s

Threshold/Upper threshold 10 mAU/ 3000 mAU



Expected result

Run a single injection under the conditions as described in Table 14. You should observe a chromatogram with four peaks as displayed in Figure 36 on page 88. Four fractions should have been collected in the fraction collector vials.

NOTE The retention times of the peaks could be different depending on the system configuration (e.g. flow cell type, capillary volumes, type and age of column).

Figure 36 Chromatogram of the check-out injection with UV-based trigger


Configuration and Operation of the Fraction Collector 2Check-out Procedures

Check-out procedure for UV-triggered fraction collection PS

Prerequisites

✔ The Agilent 1200 Series LC system has been flushed thoroughly with the liquid phase as described in Table 14 and the lamp(s) of the UV detector have been switched on for at least 15 minutes to provide a stable baseline.

✔ The Agilent 1200 Series fraction collector has been installed and configured. The delay volume has been calibrated as described in “Performing a delay calibration with an UV detector” on page 57.

✔ The 15-vial tray (G1313-44513) with at least four 6ml vials is installed.

✔ All wells are empty and the fill levels, which are stored on the module, have been reset. To verify this remove and insert the 15-vial tray. Then click on Yes in displayed dialog box.

Table 15 Method parameters for UV-triggered fraction collection check-out PS

Agilent 1200 Series modules • preparative pump plus gradient extension (optional)• preparative autosampler/ dual-loop autosampler• diode-array detector or multi-wavelength detector with

preparative flow cell*

• fraction collector preparative scale

Column ZORBAX SB-C18, 50 mm x 9.46mm (846975-202) or equivalent

Sample Check-out sample (01080-68704)

Flow 10 ml/min

Stoptime 4 minutes

Solvent A 35% (HPLC grade water)

Solvent B 65% (HPLC grade acetonitrile)

Wavelength DAD/MWD Signal A: 254/16nm, Reference: 360/80nm

Peakwidth (Responsetime) > 0.1min (2s)

Injection volume 50 µl

Column temperature ambient

Fraction trigger mode Peak-based



Expected result

Run a single injection under the conditions as described in Table 15. You should observe a chromatogram with four peaks as displayed in Figure 36 on page 88. Four fractions should have been collected in the fraction collector vials.

Max. peak duration 0.5 min

DAD/MWD working mode Threshold/slope

Upslope 5 mAU/s

Downslope 10 mAU/s

Threshold/Upper threshold 10 mAU/ 3000 mAU

* The checkout method is tailored for the 3mm stainless steel flow cell G1315-60016. For other pre-parative flow cells some parameters (e.g. flow rate, injection volume, threshold/slope), should be adjusted.

Table 15 Method parameters for UV-triggered fraction collection check-out PS

NOTE The retention times of the peaks could be different depending on the system configuration (e.g. flow cell type, capillary volumes, type and age of column).


Configuration and Operation of the Fraction Collector 2Application Notes

Application Notes

More information about the Agilent 1200 Series fraction collectors and Purification Systems are available from a series of application notes. Printed versions can be ordered from Agilent or pdf-files can be downloaded from the Agilent Website

http://www.chem.agilent.com

Table 16 Selected Agilent Technologies Application Notes

Title Publication Number

An optimized system for analytical and preparative work 5988-9649EN

Recovery collection with the Agilent LC purification system 5988-9650EN

Solutions for preparative HPLC-Application Compendium 5988-9646EN

Innovative fraction collection with the Agilent LC purification platform 5988-9250EN

Automated fraction re-analysis - does it really make sense? 5988-8653EN

Injection of high-concentration samples with the Agilent LC purification system

5988-8654EN

Sophisticated peak-based fraction collection - working with up and down slope

5988-7895EN

Strategies for purification of compounds from non-baseline separated peaks

5988-7460EN

Method scale-up from analytical to preparative scale with the Agilent LC purification system PS

5988-6979EN

Peak-based fraction collection with the Agilent LC purification system AS - Influence of delay volume on recovery

5988-5747EN


2 Configuration and Operation of the Fraction CollectorApplication Notes



3Control Module Screens of the Fraction Collector

Major keys on the Agilent 1200 Series control module 94

Screens available from the Analysis view 95

Screens available from the System view 109

This chapter is intended to introduce an operator to the functionality available for operation of the Agilent 1200 Series fraction collector with the Agilent 1200 Series control module.

Please use the manual of Control module for further detailed reference.


3 Control Module Screens of the Fraction CollectorMajor keys on the Agilent 1200 Series control module

Major keys on the Agilent 1200 Series control module

ESC Return to previous screen, abort any change of parameters and toggle between the last two top layer views

m Open context sensitive menus

i Information/Help

Enter Store changed parameters or execute a choice

Done (If available) Activate settings of current screen

On/Off Switch on individual Instrument(s) or complete System

Start Start a location range or sequence

Plot View online signals

Views Change between analysis - (samples)- status - system views

NOTE The screens shown on the next pages are based on:Control Module (G1323B) firmware revision B.03.0x and higher LC Module firmware revision A.05.06 and higher

NOTE In case the control module’s display seems to be frozen (hang-up due to a communication problem on the CAN bus), unplug the control module from the HPLC module and reconnect.


Control Module Screens of the Fraction Collector 3Screens available from the Analysis view

Screens available from the Analysis view

Analysis view This is the wake-up screen, if the Agilent 1200 Series fraction collector or thermostatted fraction collector is the only configured Agilent 1200 Series LC module.

On/Off (F7) allows you to turn on the Thermostat Module of the thermostatted versions of the fraction collector.

The m-key allows access to the context sensitive menus. Date&Time allows you to change time settings. Print Screen gives access to the print screen. About tells you the current firmware revision and the serial# of your control module. Setup view leads you to analysis view configuration for additional Agilent 1200 Series LC modules. Restart re-boots the control module. If multiple instruments are configured in the Analysis screen the context menu contents will vary, depending on the cursor position, when the m-key is pressed (depending on highlighted module).


3 Control Module Screens of the Fraction CollectorScreens available from the Analysis view

In the Setup view, modules can be added or removed to the analysis view. Select a module, you want to move, with the help of the selection-keys. Use function keys F7/F8 (Remove/Add) to move the highlighted module. Changes must be activated with Done (F6).

Here, e.g. the binary pump, the injector and the detector parameters are shown on the display as well. The number of parameters for each module is reduced as additional modules are added to the display. A maximum of 4 modules can be shown simultaneously. If more modules are connected to the system, you have to choose 4 of them in Setup view.



Settings With the F1-key (Settings) you open a pull-down menu where you can select the fraction collector module.

Within Settings you can change the fraction collector parameters like the type of fraction collection (Off, Timetable, or Peak Controlled). There are different sets of parameters available through the F1-5 navigation-keys for setting path operation of the fraction collector. F7 (Default) resets the fraction collector to default values. Changes must be acknowledged with F6 (Done).

Settings screen with thermostat connected. On/Off (F8) allows you to switch the thermostat on or off.



Use the m-key for the context sensitive menu. The Status command pulls up a module specific screen displaying details of configured trays. Reset will initialize the fraction collector.

Settings More... The F1-key (More) opens a pull-down menu.

By selecting More...Auxiliary on the pull-down menu you can enter the fraction collector Fill Volume Per Vessel for the currently installed trays and vessels. Leaving this value at the “DEF” value will completely fill the vessels according to their specified size. Changes must be acknowledged with F6 (Done).



By selecting More...Rinse Needle on the pull-down menu you can specify when the needle is rinsed. Settings can be Off (both tick marks not selected), At Start of Analysis or Between Fractions. Rinsing the needle between fractions is only available, for peak-based fraction collection with the analytical scale fraction collector G1364C. Changes must be acknowledged with F6 (Done).

By selecting More...Flow on the pull-down menu you can specify the flow rate of the connected pump, in case the pump is a non-1200 Series pump or if the pump is not connected to the fraction collector via CAN.



Settings -Timetable

With the F2-key (Timetable) in the Settings screen you can list the timetable for the fraction collector. Press F7-key (Insert) to edit the selected entry or F6-key (Delete) to remove the selected/highlighted entry.

Edit each entry line according to your needs, by entering a time for a Setting and by selecting a Setting with the help of the pull-down menu. Use F7 (Enter) to insert the current settings. Changes must be acknowledged with F6 (Done).



Use the m-key for the context sensitive menu. It gives you additional tools for the timetable.

In the timetable several trigger modes can be selected. Peak-based trigger can be specified for defined time frames during the run. For time-based trigger either the duration of a fraction or the number of fractions per time interval can be chosen.



Settings -Detectors

With the F3-key (Detectors) in the Settings screen you can configure and Enable (F8) or Disable (F7) any detector that is part of a system for peak detection. Changes must be acknowledged with F6 (Done).

Settings -Thermostat

With the F4-key (Thermostat) in the Settings screen you can set the fraction collector thermostat temperature. Changes must be acknowledged with F6 (Done).



Settings -Run times

With the F5-key (Runtimes) in the Settings screen you can change the stop time and the post-run time for the selected module individually. Changes must be acknowledged with F6 (Done).

Method screens Use Esc to return to the Analysis screen. Use the F3-key (Method) to view the parameters in a method. Use the F8-key (Save As) to save the method in the module(s). The PC-Card-key is only active when a PCMCIA card is inserted in the control module. Module (F1) and Method (F2) give access to methods stored on the Agilent 1200Series LC module or the PC-Card. Timetable (F3) gives access to the timetable of a method (if specified).

NOTE The PCMCIA card must have been inserted into the control module prior to the last start-up in order to be recognized.



After pressing F1 or F2 use the Right/Left selection keys to switch between PC-Card and Instrument window.Use the Up/Down selection keys to select the method. Use F7/F8 (Copy) to copy a selected method from one window to the other one. Use F6 (Delete) to delete a selected method.

Press the m-key to open the context sensitive menu for printing the method directory.



sequence In the sequence view (F4 from the Analysis Screen) you can specify the sequence’s End Actions (F1), load sequences from PC-Card (F2) (if present) or proceed to a method table (F3). F6 (Delete) and F7 (Insert) allow you to edit lines in the sequence table.

Parameters that can be entered into a line in the sequence table.



Fractions From the Analysis View press the F5-key (Views) and select Fractions.

This is an example if an Agilent 1200 Series fraction collector is configured stand-alone. Here you can define a Start position for a fraction, select a collection Scheme, choose a Trigger Mode etc.

status From the Analysis View press the F5-key (Views) and select Status.



This is an example if an Agilent 1200 fraction collector is configured stand-alone.

Signal plot Press F6-key (Plot) to enter the plot screen (available also from all other screens). Here you can observe the on-line signal(s). If more than one signal is configured, use the 1-2-3 alphanumeric keys to switch between the signals.

Press the m-key to open the context sensitive menu.



Press F6-key (Select). Here you can add additional on-line signals (maximum are 3). Additional signals could be also pressure or temperature signals from other modules. Use the Right/Left selection keys to switch between ‘Available Signals’ and ‘Selected Signals’. Use the F8-key (Move) to enter available signals into the box for selected signals or vice versa. Use the F7-key (Setup) to enter the display range for the highlighted/selected signal. Use F6 (Done) to activate changes and to return to the Plot screen.


Control Module Screens of the Fraction Collector 3Screens available from the System view

Screens available from the System view

System view Use the Esc-key until you receive Views on the F5-key. Choose System from the pull-down menu. This screen shows the last activities in the system.

System - Control Use the F1-key (Control) to select the fraction collector. Here you receive information about the not-Ready conditions, if needed. Press F2 to Home the transport unit (e.g. for replacing trays), press F3 to Park the carrier for transportation (also requires a protective foam and cardboard holder to avoid damage to the transport unit during transportation). F7 (Reset) does a re-initialization of the fraction collector. Use the F8-key (On/Off) to turn on the thermostat (if present). Changes must be acknowledged with F6 (Done).


3 Control Module Screens of the Fraction CollectorScreens available from the System view

Manual Trigger In this screen you can view the online signal of any connected detector. Press F6 (Select) to choose, which detector signal will be displayed for identifying the starting point and the end point of a fraction. The Manual Trigger Mode allows to manually start a fraction collection by pressing 1-Trigger (F8). Pressing 0-Trigger, again, will then stop the collection of a fraction. The fraction collector will wait for the delay-time, specified for the chosen detector, before it starts or stops the collection of a fraction.



System -Configure

Use the Esc-key to receive Views on the F5-key in the Analysis Screen. Choose System from the pull-down menu. Use the F2-key (Configure) to select the fraction collector. Here you define further special set points for the fraction collector operation. Changes must be acknowledged with F6 (Done).

Use the F1-key (Interfaces) to access the interface settings (if required). Changes must be acknowledged with F6 (Done).



Configure - trays Use the F2-key (Trays) to view and configure the (automatically) detected sample trays. Different views will come up depending on the installed trays. Enter the Vessel Volume and the Vessel Height for the vessels used with your installed tray(s). Changes must be acknowledged with F6 (Done).

Select/highlight a tray configuration with the help of the selection keys. F7/F8 (Add/Remove) allows you to move configurations from available Plate Formats to the Active Plates and vice versa. Press F2 (Delete Def.) to delete a user defined definition. Press F3 (Rotate Plate) to rotate the selected plate. Use F1 (Definition) to edit the user defined definition of a well-plate. Changes must be acknowledged with F6 (Done).



Configure -fraction

F3 (Fraction) in the main Configuration screen allows you to access the Delay volume definition screen, to specify Filling Order and Reserved or Recovery Locations.

Configure -fraction - delays

Delays in the Fraction Configuration screen allows you to configure the delay volume between each connected detector and the fraction collector. Edit (F8) allows to enter new values for the selected detector. Changes must be acknowledged with F6 (Done).



Configure -fraction -

filling order

Filling Order in the Fraction Configuration screen allows you to configure the filling order of the trays in the fraction collector. Edit (F8) allows to enter new values for the selected detector. Changes must be acknowledged with F6 (Done)

Configure -reservedlocations

Reserved Locations allows you to specify reserved locations by Rows, Columns or Single Locations, that are not used (filled) during the collection. Changes must be acknowledged with Done (F6).



Configure -recovery location

The Recovery Location allows you to specify a location for the collection of any mobile phase or sample that are not collected with the fractions during the study. Changes must be acknowledged with Done (F6).

Configure -Collection Mode

In the Collection Mode View it can be specified if the valve switches to waste when the needle moves to the next vessel or if the needle Moves without Valve Switching.

Configure -Needle

Movement

The Needle Movement allows you to specify the position of the needle tip during fraction collection. With into location the needle tip moves to the specified depth in the well. With above location the needle tip stays at the specified distance above the well. In the contact control mode the needle tip moves to the specified distance above the bottom of the well. When the fraction is collected the needle will move upwards while staying in contact with the liquid surface of the collected fraction. This movement types enables the Agilent 1200 Series fraction collector to collect low volumes without carry-over or air bubbles at the bottom of the well. Changes must be acknowledged with Done (F6).



Configure -Flow Path

In the Flow Path configuration the diameter fraction collector capillary and the Needle Type can be defined.

Configure -Thermostat

F4 (Therm) in the main Configuration screen allows you to configure the temperature and turn on conditions of the thermostatted fraction collector. Changes must be acknowledged with F6 (Done).



Configure -Needle move

Press F5 from the Configure Screen and choose how the fraction collector’s needle carrier arm moves from one location to the next one. This can either be with diverting the flow to waste (with valve switching) while moving (default) or without diverting the flow (without valve switching, optional only when neighbored well plate positions are used). With the analytical scale fraction collector it is also possible to insert the needle into a capped vial or through a closing mat of a well plate with a specified depth (Move into vessel).

System - tests Press F3 from the System View and choose the fraction collector to access the fraction collector Tests screen.

The Tests screen gives you access to multiple tests and features such as the automatic transport unit Alignment (F1) tool, maintenance procedures to Change (F2) parts on the fraction collector or additional test Signals (F3) to monitor various parameters during the fraction collector operation. Calibration (F4) gives you access to the delay calibration procedure. Parts Info (F5) gives you information about the type and revision of the installed transport unit assembly. The pull-down menu in the center of the screen gives you access to various step commands.



Use the F1-key (Align Trans) to perform an automatic alignment of the fraction collector transport unit.

The alignment process may take several minutes. Changes must be acknowledged with F6 (Done).

EMF (EarlyMaintenance

Feedback)

Use the F1-key (EMF) to set EMF limits. Choose Setup limits to select the number of injection valve cycles or number of needle movements into the needle seat at which you want to receive a warning. Changes must be acknowledged with F6 (Done).

WARNING Only the empty 4-plate tray base (G1364-84501) should be installed, when performing the alignment. The well plates must be removed from the fraction collector tray base before performing the transport unit alignment. If the well plates are installed during the alignment procedure, the transport unit might be damaged.



If a set limit has been exceeded, a message box will pop up. If you press Reset, the limits will be removed. Ignore will continue to keep the EMF flag set.

NOTE An exceeded limit will not stop a sequence or run (information only, to plan maintenance activities).



System /Error Log

Use the F2-key (System Log) or F3-key (Error Log) to look for errors. For troubleshooting reasons these Logbooks can be printed (F6) or saved to a file on the PCMCIA card (pressing the m-key).



Maintenance Log Use the F4-key (Maintenance Log) to look for maintenance activities performed on the instrument. A list of possible events are listed in the scroll screen (Needle replaced, Inlet/waste tubing replaced, Valve-to-needle tubing replaced, Funnel seals replaced, Rinse port seal replaced). Select the activity you have performed and press Done to create an entry in the logbook. Editing the add line with the help of the alphanumeric keys allows you to enter your own maintenance function. For troubleshooting reasons this Logbook can be printed or saved to a file on the PCMCIA card (pressing the m-key). Changes must be acknowledged with Done (F6).



Firmware Update Use the Esc-key to receive Views on the F5-key. Choose System from the pull-down menu. Use the F3-key (Records) to select the fraction collector. Use the F5-key (FW Update) to enter the Update section. If you want to update the resident firmware (together with specific main firmware revisions), select the a file from the PCMCIA card (RESnnnn.DLB) and press execute. If you want to update the main firmware, press F7-key (Transfer) to turn the module into the resident mode (LED on module should blink yellow).

Use the Esc-key to receive Views on the F5-key. Choose System from the pull-down menu. Use the F3-key (Records) to select the fraction collector. Use the F5-key (FW Update) to enter the Update section. Select the a file from the PCMCIA card (1364nnnn.DLB for the fraction collector) and press execute. When the update has finished the update, press F8-key (Transfer) to return the module into the normal mode (LED on module should stay yellow).



Use the F5-key (FW Update) to enter the Update section. Select the a file from the PCMCIA card (1364nnnn.DLB for the fraction collector) and press execute. When the update has finished, press F7-key (Transfer) to return the module into the normal mode (LED on module should stay yellow).

If you have not saved your methods, please do it before continuing. Otherwise they will be overwritten during the update process.



Changing theserial number

In case the serial number of the module has to be added, use the m-key to open the menu Enter Serial#. The serial number becomes active after restart of the module.



Type in the module’s Serial# as indicated on the front or rear label of the module using the alphanumeric keys. Changes must be acknowledged with F6 (Done).



4Troubleshooting and Test Functions

Status Indicators 129



Transport Unit Self Alignment 133

Step Commands 135


4 Troubleshooting and Test Functions

Status Indicators

The fraction collector is provided with two status indicators which indicate the operational state (prerun, not ready, run, and error states) of the instrument. The status indicators provide a quick visual check of the operation of the fraction collector (see “Status Indicators” on page 129).

Error Messages

In the event of an electronic, mechanical or hydraulic failure, the instrument generates an error message in the user interface. For a detailed description of the failure, a list of probable causes of the problem, and a list of suggested actions refer to the Agilent 1200 Series Fraction Collectors Service Manual G1364-90110.

Maintenance Functions

The maintenance functions position the transport unit and needle carrier assembly on certain positions for maintenance, homing, parking or delay calibration (see “Maintenance Functions” on page 131).

Transport Unit Self Alignment

The transport unit alignment with the transport unit and the well-plate tray is required to compensate for larger deviations in positioning the needle carrier assembly.

The transport unit self alignment is required after disassembling the system or when you exchange the transport unit, the needle carrier assembly or the MTP main board. This function is in the Diagnose screen of the ChemStation or the Control Module.

WARNING The sample transport self alignment requires one of the 4-well-plate trays (Part Number: G1364-84501), but well plates MUST NOT be installed!


Troubleshooting and Test Functions 4Status Indicators

Status Indicators

Two status indicators are located on the front of the fraction collector. The lower left indicates the power supply status, the upper right indicates the fraction collector status.

Power Supply Indicator

The power supply indicator is integrated into the main power switch. When the indicator is illuminated (green) the power is ON.

Figure 37 Location of Status Indicators

Status indicatorgreen/yellow/red

Line power switch with green light


4 Troubleshooting and Test FunctionsStatus Indicators

Instrument Status Indicator

The instrument status indicator indicates one of four possible instrument conditions:

• When the status indicator is OFF (and power switch light is on), the instrument is in a prerun condition, and is ready to begin an analysis.

• A green status indicator indicates the instrument is performing an analysis (run mode).

• A yellow status indicator indicates a not-ready condition. The instrument is in a not-ready state when it is waiting for a specific condition to be reached or completed (for example, front door not closed), or while a self-test procedure is running.

• An error condition is indicated when the status indicator is red. An error condition indicates the instrument has detected an internal problem which affects correct operation of the instrument. Usually, an error condition requires attention (for example, leak, defective internal components). An error condition always interrupts the analysis.


Troubleshooting and Test Functions 4Maintenance Functions

Maintenance Functions

Some maintenance procedures require the needle arm, and needle carrier to be moved to specific positions to enable easy access to components. The maintenance functions move these assemblies into the appropriate maintenance position. In the ChemStation the fraction collector maintenance positions can be selected from the Maintenance menu in the Diagnosis view (see Figure 38). In the Control Module the functions can be selected in the Test screens of the fraction collector.

Change Parts.

If you click Start the transport unit will move upwards, the needle carrier assembly will move to the front center and then turn OFF the theta motor to allow free rotation of the theta arm. This position enables easy access to the transport unit to change:

Figure 38 Fraction Collector Maintenance Positions dialog box


4 Troubleshooting and Test FunctionsMaintenance Functions

• The needle assembly

• The needle carrier assembly

• The valve to needle tubing.

• The diverter valve.

After the maintenance or repair task has been finished, click End to move the transport assembly to the Home position

Home Position

This maintenance function moves the arm up and to the right rear for better access and exchange of the trays.

Park Arm

This maintenance position moves the arm to the park position at the upper rear left side of the tray for transporting or shipping the fraction collector.


Troubleshooting and Test Functions 4Transport Unit Self Alignment

Transport Unit Self Alignment

The transport unit alignment with the transport unit and the well-plate tray is required to compensate for larger deviations in positioning the needle carrier assembly.

The transport unit self alignment is required after disassembling the system or when you exchange the transport unit, the needle carrier assembly or the MTP main board.

This function is in the Diagnose screen of the ChemStation or the Control Module.

When is a Transport Unit Self Alignment Necessary?

The sample transport self alignment is required after disassembling the module or when you exchange:

• The transport unit.

• The needle/capillary carrier assembly.

• The MTP main board.

WARNING The sample transport self alignment requires one of the 4-well-plate trays (Part Number: G1364-84501), but well plates MUST NOT be installed.

If the Transport Unit Self Alignment is started with well plates on the tray, the alignment procedure is aborted WITHOUT error message.

How to perform a Transport Unit Self Alignment?

Steps Comments

1 If the transport unit has been exchanged or if it is very badly misaligned, set the 8-bit configuration switch to the Forced Cold Start Configuration.

For details see: “Forced Cold Start” in the Fraction Collectors Service Manual.

2 Install the 4-well-plate tray (G1364-84501) IMPORTANT: Remove all plates!


4 Troubleshooting and Test FunctionsTransport Unit Self Alignment

3 Ensure that the wellplate adapter is correctly assembled

On a G1364B preparative scale fraction collector install the delay calibration adapter G1364-87301

4 Select the Maintenance menu in the Diagnosis view of the Agilent ChemStation.

5 In the menu choose Fraction Collector > Transport Alignment... to start the automated procedure.

The Transport Alignment Procedure takes approximately 10-15 minutes

6 Set the 8-bit configuration switch to the default setting.

See “8-bit configuration switch” in the Fraction Collectors Service Manual.

How to perform a Transport Unit Self Alignment?

Steps Comments

NOTE If the Transport Unit Self Alignment is started with well plates on the tray, the alignment procedure is aborted without any error message.


Troubleshooting and Test Functions 4Step Commands

Step Commands

Some movements of the fraction collection sequence can be done under Manual control. This is useful during troubleshooting where close observation of each of the fraction collection step is required to confirm a specific failure mode or verify successful completion of a repair.

Each step command actually consists of a series of individual commands which move the fraction collector components to predefined positions enabling the specific step to be done.

In the ChemStation the step commands can be selected from the “Test Selection Box” (see Figure 39) in the Diagnosis display. In the Control Module the step commands can be accessed from the pull-down menu in the fraction collector “Test”.

Figure 39 Fraction Collector Step Commands


4 Troubleshooting and Test FunctionsStep Commands

Troubleshooting

If the fraction collector is unable to perform a specific step due to a hardware failure, an error message is generated. You can use the step commands to perform a fraction collection sequence, and observe how the fraction collector responds to each command.

Table 17 Step Commands

Step Action Comments

Needle Up Lifts the needle arm to the upper position.

Command also switches the diverter valve to waste if it is not already in that position.

Needle into vessel Lowers the needle into the specified vessel.

only analytical scale G1364C

Needle to rinse / flush port

Moves the needle to the rinse / flush port.

only analytical scale G1364C

Switch valve to needle Switches the diverter valve to the needle.

Switch valve to waste Switches the diverter valve from needle to waste.



5Repairing the Fraction Collector

Cleaning the Fraction Collector 138

Overview of Main Repair Procedures 139

Simple Repairs 140


5 Repairing the Fraction CollectorIntroduction into Repairing the Fraction Collector

Introduction into Repairing the Fraction Collector

Simple Repairs

The fraction collector is designed for easy repair. The most frequent repairs such as changing a needle assembly or tubings can be done from the front of the instrument with the instrument in place in the system stack. These repairs are described in “Simple Repairs” on page 140.

Cleaning the Fraction Collector

The fraction collector covers should be kept clean. Cleaning should be done with a soft cloth slightly dampened with water or a solution of water and a mild detergent. Do not use an excessively damp cloth from which liquid could drip into the fraction collector.

WARNING When opening capillary or tube fittings solvents may leak out. Please observe appropriate safety procedures (for example, goggles, safety gloves and protective clothing) as described in the material handling and safety data sheet supplied by the solvent vendor, especially when toxic or hazardous solvents are used.

WARNING To avoid personal injury, keep fingers away from the needle area during fraction collector operation. Do not bend the safety flap away from its position, or attempt to insert or remove a vial from the gripper when the gripper is positioned below the needle.

WARNING Regularly inspect the inlet / waste tubing assembly, and the valve to needle tubing and exchange them if they are worn out or show visible signs of damage.

WARNING Do not let liquid drip into the fraction collector. It could cause a shock hazard or damage to the fraction collector.


Repairing the Fraction Collector 5Overview of Main Repair Procedures

Overview of Main Repair Procedures

Figure 40 Main Assemblies (Example displays the analytical scale module G1364C. The preparative scale fraction collector G1364B contains a different needle)

Internal tray withrinse port and flow

delay sensor

Diverter Valve

Transport unitBCD Board (optional)

Main board

Needle

Illumination Assembly

Needle carrier

Leak plane withleak sensor

Tray compartmentdivider

Ribbon cable

Inlet / waste tubings

Wellplate Adapter

Plug channel with air channel adapter underneath


5 Repairing the Fraction CollectorSimple Repairs

Simple Repairs

The procedures described in this section can be done with the fraction collector in place in the stack. These procedures can be done on a more frequent basis.

Table 18 Simple Repair Procedures

Procedure Typical Frequency Notes

Replacing the Inlet / waste tubings

When worn out, when showing visual signs of damage, typically once per year.

See “Replacing the Inlet/Waste Tubings” on page 141

Replacing the valve to needle tubing

When worn out, when showing visual signs of damage, typically once per year

See “Replacing the Valve to Needle Tubing” on page 145

Exchanging the preparative needle assembly

When needle shows indication of damage or blockage

See “Exchanging the Preparative Needle Assembly” on page 148

Exchanging the analytical needle assembly

When needle shows indication of damage or blockageOr when using the short needle assembly for operation with high test tubes (>45mm)

See “Exchanging the Analytical Needle Assembly” on page 150

Exchanging the diverter valve When defective (internal / external leak, valve not switching any more)

See “Exchanging the Diverter Valve” on page 153

Exchanging the internal tray When flow delay sensor defective See “Exchanging the Internal Tray” on page 156

Repairing or exchanging a funnel of the internal tray or funnel tray

When defective (leaky, blocked or contaminated)

See “Repairing or Exchanging a Funnel of the Internal Tray” on page 158


Repairing the Fraction Collector 5Simple Repairs

Replacing the Inlet/Waste Tubings

Frequency When contaminated, worn out or visibly damagedTypically once every year

Tools required None

Parts Required Inlet / waste tubing assembly in tubing kit preparative scale 0.8 mm ID, PN G1364-68711, analytical scale 0.25 mm ID, PN G1364-68712 or analytical scale 0.15 mm ID, G1364-68723

WARNING To avoid personal injury, keep fingers away from the needle area during fraction collector operation.

WARNING Follow the described installation procedure exactly to maximize the lifetime of the inlet / waste tubings and to avoid potential spills or fraction losses. Regularly inspect the tubings and exchange them if they are worn out or show visible signs of damage.



Before beginning this procedure:

• Position the transport unit of the fraction collector in the “Home Position” (see “Maintenance Functions” on page 131).

• Remove all installed trays from the tray base.

• Position the transport unit of the fraction collector in the “Change Parts Position” (see “Maintenance Functions” on page 131).

• Turn OFF the instrument.

• Remove the rear end of the fraction collector’s waste tubing from the waste container, unscrew the front end of the fraction collector’s inlet tubing from the flow cell of the detector.

1 Locate the diverter valve with the finger-tight fittings of the inlet / waste tubing assembly (the figure shows the open fraction collector seen from the front).

2 Unscrew the 2 finger-tight fittings of the inlet / waste tubing assembly at the diverter valve.

3 Unplug the inlet / waste tubing assembly from the bolt carrier (located on the top center of the front cover) by moving the snapper as indicated by the arrows. Slide the tubings out to the bottom.

1

2



4 Install the new inlet / waste tubing assembly into the bolt carrier as shown below. Slide in the long ends of the tubings from bottom to top and let the snapper click into position.

5 The rear tubing (with the label with the arrow) must be connected to the flow cell of the detector. The front tubing must be inserted into the waste container.

6 Connect the finger-tight fittings of the inlet / waste tubing assembly to the ports of the diverter valve. IMPORTANT: The tubings must not be bent up- or downwards. The cables must not be twisted. (View from the bottom)

7 A color coded ring on one of the tubings and the valve body indicates, which cable belongs to which port.IMPORTANT: It is absolutely vital to connect these tubings as described, in order to maximize their lifetime and operating security.

From flow cellTo waste



8 IMPORTANT: The cables must run into the ports of the diverter valve in lines parallel to the horizon.

On completion of this procedure:

• Re-install the tray(s) in the tray base.

• Start the instrument.

• Close the front cover.



Replacing the Valve to Needle Tubing

Frequency When contaminated, worn out or visibly damagedTypically once every year

Tools required Wrench, open end, 4mm, PN 8710-1534 (supplied in accessory kit)Wrench, open end, 1/4 – 5/16 inch, PN 8710-0510 (supplied in accessory kit)

Parts Required Valve to needle tubing assembly included in tubing kit preparative scale 0.8 mm ID, PN G1364-68711, analytical scale 0.25 mm ID, PN G1364-68712 or analytical scale 0.15 mm ID, G1364-68723


WARNING Follow the described installation procedure exactly to maximize the lifetime of the valve to needle tubing and to avoid potential spills or fraction losses. Regularly inspect the tubings and exchange them if they are worn out or show visible signs of damage.




• Position the transport unit of the fraction collector in the “Change Parts Position” (see “Maintenance Functions” on page 131) and turn off the instrument.

• It might be more convenient to remove the needle from its carrier before unscrewing the needle tubing.

1 Locate the diverter valve with the finger-tight fittings of the valve to needle tubing assembly (the figure shows the open fraction collector seen from the front).



2 Unscrew the finger-tight fitting of the valve to needle tubing assembly at the diverter valve.

3 Using the 4 mm wrench and the 5/16“ wrench for counter-holding unscrew the valve to needle tubing from the needle. (Viewed from the bottom)

4 Un-clip the tubing from the bottom of the needle carrier assembly.

5 Slide the tubing through the hole in the needle carrier assembly (from bottom to top) and out of the holder in the z-arm assembly.

1

2



6 Install the new valve to needle tubing assembly by clipping it in to the holder in the z-arm assembly (1), Important!!!) and slide it through the hole in the z-arm (2) and out on the bottom of the needle carrier assembly (top to bottom). Screw the finger-tight fitting into the port of the diverter valve (3).

7 Using the 4 mm wrench and the 5/16“ wrench for counter-holding connect the valve to needle tubing to the needle. (Viewed from the bottom)

8 IMPORTANT: After fixing the screw clip the tubing into the guide on the bottom of the needle carrier assembly.It is absolutely vital that the tubing is installed as described, to maximize the lifetime of the tubing.


• Re-install the needle to the needle carrier assembly, if you previously removed it. Make sure to slide the needle all the way to the front of the needle carrier assembly (clicks into position).




2

1

3



Exchanging the Preparative Needle Assembly

Frequency When the needle is leaky or visibly damagedWhen the needle is blocked or contaminated


Parts required Preparative needle assembly, G1364-87201


CAUTION Regularly inspect the inlet / waste tubing assembly and the valve to needle tubing and exchange them if they are worn out or show visible signs of damage.







1 Using the 4 mm wrench and the 5/16“ wrench for counter-holding unscrew the valve to needle tubing from the needle. (Viewed from the bottom)



2 Holding the needle assembly between your thumb and forefinger, slide out the assembly towards the rear of the needle carrier assembly.

3 Insert the new needle assembly into the holder of the needle carrier assembly. Make sure to push it all the way to the front.

4 Using the 4 mm wrench and the 5/16“ wrench for counter-holding connect the valve to needle tubing to the needle. (Viewed from the bottom)


• Re-install the needle to the needle carrier assembly, if you previously removed it. Make sure to slide the needle all the way to the front of the needle carrier assembly (clicks into position).

• IMPORTANT: After fixing the needle in the needle carrier clip the tubing into the guide on the bottom of the needle carrier assembly in case it slipped out of this guide (see 2 left arrows on the figure to the left).It is absolutely vital that the tubing is installed as described, to maximize the lifetime of the tubing.






Exchanging the Analytical Needle Assembly

Frequency When the needle is visibly damagedWhen the needle is blocked or contaminated


Parts required Analytical needle assembly, G1367-87200


CAUTION Regularly inspect the inlet / waste tubing assembly and the valve to needle tubing and exchange them if they are worn out or show visible signs of damage.







1 Use the 4 mm wrench and the 5/16” wrench for counter-holding to unscrew the valve-to-needle tubing from the needle assembly. (Viewed from the bottom)



2 Remove the wellplate adapter by turning it clockwise (1). Hold the needle assembly between your thumb and forefinger, slide out the assembly towards the rear of the needle carrier assembly (2).

3 Insert the new needle assembly into the holder of the needle carrier assembly. Make sure to push it all the way to the front. Reassemble the wellplate adapter.

4 Use the 4 mm wrench and the 5/16“ wrench for counter-holding to connect the valve to needle tubing to the needle. Reassembly the wellplate adapter. (Viewed from the bottom)


• IMPORTANT: After fixing the needle in the needle carrier clip the tubing into the guide on the bottom of the needle carrier assembly in case it slipped out of this guide (see 2 left arrows on the figure to the left).It is absolutely vital that the tubing is installed as described, to maximize the lifetime of the tubing.




1.

2.



Exchanging the Diverter Valve

Frequency When leaky or defective

Tools required Hex key 2.0 mm, PN 8710-2438 (supplied in accessory kit)

Parts required Diverter Valve, PN G1364-61901






1 Locate the diverter valve with the finger tight fittings of the inlet / waste tubing assembly (the figure shows the open fraction collector seen from the front).

2 Unscrew the 2 finger tight fittings of the inlet / waste tubing assembly at the diverter valve.

3 Unscrew the finger tight fitting of the valve to needle tubing assembly at the diverter valve.



4 Disconnect the power cable of the diverter valve from the connector on the front of the z-arm.

5 Using the 2 mm hex key loosen the holding screw of the diverter valve and remove the diverter valve from the z-arm.

6 Install the new diverter valve to the z-arm with its holding screw. Plug in the new diverter valve power cable to the z-arm.

7 Install the valve to needle tubing at the new divertervalve.

2

1



8 Connect the finger tight fittings of the inlet / waste tubing assembly to the ports of the diverter valve. IMPORTANT: The tubings must not be bent up- or downwards. The cables must not be twisted. (View from the bottom)

9 A color coded ring on one of the tubings and the valve body indicates, which cable belongs to which port.IMPORTANT: It is absolutely vital to connect these tubings as described, in order to maximize their lifetime and operating security.

10 IMPORTANT: The cables must run into the ports of the diverter valve in lines parallel to the horizon.







Exchanging the Internal Tray

Frequency When defective

Tools required None

Parts required Internal tray analytical scale, (G1364-63113)Internal tray preparative scale,(G1364-63114)





1 Locate the internal tray assembly with the rinse funnel and flow delay sensor in the bottom of the right front corner of the instrument.



2 Remove the internal tray by pushing down the plastic holder that holds it in position underneath the metal latch (1) and sliding the tray to the left at the same time (2).

3 Remove the corrugated waste tubing from the front of the instrument (1) and slide the internal tray’s waste tubing to the rear of the outlet (2) the before sliding the tray out.

4 Install the new tray by sliding it into position underneath the metal latch that holds it. The waste tubing from the internal tray should be guided as shown.

5 Make sure that the waste tubing is slid all the way through the outlet (1). Its end should be over the edge and below the level of the laboratory desk that the system stands on to avoid any back flow of solvent. Re-install the corrugated waste tubing (2).





2

1 CellFrame

12

CellFrame

2

1



Repairing or Exchanging a Funnel of the Internal TrayFrequency When leaky or contaminated

Tools required None

Parts required Funnel assembly, waste tubing assembly, seals (for part numbers see “Internal Tray Assembly” on page 176)


• Position the transport unit of the fraction collector in the “Home Position” (see “Maintenance Functions” on page 131) and remove all installed trays from the tray base.


• The procedure is almost identical for both, the preparative and the analytical scale internal tray, but it requires different parts.

1 Remove the internal tray from the instrument, see “Exchanging the Internal Tray” on page 156.

2 Turn the internal tray upside down and remove the funnel’s waste tubing through the flow delay sensor.

3 Remove the funnel assembly from the internal tray by screwing it counter clockwise and lifting it out.

2

1



4 Exploded view for the parts of the internal tray. Once the funnel has been removed from the internal tray, it can be disassembled and defective parts can be replaced (funnel screw (1), seals (2), tubing (3) or funnel holder (4)).

5 Slide the waste tubing of the newly assembled funnel assembly through the slit beside the funnel in the internal tray assembly

6 Turn the internal tray upside down. Slide the funnel’s waste tubing through the flow delay sensor.

7 Turn the internal tray upside down.Slide the funnel’s waste tubing through the flow delay sensor.

1

2

3

4



8 Insert the funnel into the tray and screw it tight (clockwise)

9 Re-install the plug that holds the flow delay sensor (1), pull the waste tubing tight through the flow delay sensor (2) and mount the waste tubing (3).

10 Re-install the internal tray and make sure to route the waste tubing out through the outlet of the instrument, correctly (see “Exchanging the Internal Tray” on page 156).





1

3

2



6Parts and Materials

Supported Trays for Fraction Collectors 162

List of Recommended Test Tubes 164

List of Recommended Vials and Caps 165

List of Recommended Plates and Closing Mats 168

Transport Unit Assembly (Preparative Scale) 170

Transport Unit Assembly (Analytical Scale) 171

Needle Assemblies 173

Diverter-Valve Assembly 174

Tubing Kits 175

Internal Tray Assembly 176

Fraction Collector Accessory Kit 177


6 Parts and MaterialsSupported Trays for Fraction Collectors

Supported Trays for Fraction Collectors

Table 19 Trays for the Fraction Collector

Item Description Part Number

1 Full tray for 215 test tubes, 12x100 mm G1364-84516

2 Full tray for 126 test tubes, 16x100 mm G1364-84525

3 Full tray for 60 round bottom tubes, 25x100 mm G1364-84524

4 Full tray for 40 round bottom tubes, 30x100 mm G1364-84523

5 Full tray for 4 well plates G1364-84521

6 Std. tray for 2 well plates + 10 2ml vials G2258-60011

7 Std. tray for 2 well plates + 10 collecting funnels G1364-84522

8 Funnel for item 7 5022-2200

9 Funnel seal kit for item 7 (pack. of 10) G1364-68730

10 Tubing kit 10T for item 7 (pack of 10) G1364-86707

11 Funnel coupler for item 7 G1364-43201

12 Std. tray for 100 x 2 ml vialsStd. tray for 100 x 2 ml vials, thermostattable

G1313-44510G1329-60011

13 Half tray for 15 x 6 ml vials G1313-44513

14 Half tray for 40 x 2 ml vials G1313-44512

15 Adapter air channel (installed underneath plug channel, if the fraction collector is used with the thermostat)

G1329-43200

16 Plug channel G1364-47200

17 Half Tray for 40 Funnels G1364-84532

NOTE Only one type of well-plates can be used at a time in one tray.


Parts and Materials 6Supported Trays for Fraction Collectors

Figure 41 Trays

6,7

5

43

12

1314

15 16

21

The items in this figure are displayed in different scaling factors!

17

89

10

11


6 Parts and MaterialsList of Recommended Test Tubes

List of Recommended Test Tubes

Table 20 Round Bottom Test Tubes

outer diameter height Volume recommended tray type Part Number

30 mm 100 mm 45 ml G1364-84503 (40 tubes) 5042-6459 (100/pk)

30 mm 48 mm 20 ml; G1364-84503 (40 tubes) 5042-6458 (100/pk)

25 mm 100 mm 35 ml G1364-84504 (60 tubes) 5042-6470 (100/pk)

16 mm 100 mm 19 ml G1364-84505 (126 tubes) 5022-6532 (250/pk)

16 mm 48 mm 9 ml G1364-84505 (126 tubes) 5022-6533 (100/pk)

12 mm 100 mm 11 ml G1364-84506 (215 tubes) 5022-6431 (250/pk)

12 mm 48 mm 5 ml G1364-84506 (215 tubes) 5022-6435 (100/pk)


Parts and Materials 6List of Recommended Vials and Caps

List of Recommended Vials and Caps

Table 21 Crimp Top Vials (Caps for Use with the Analytical Scale Fraction Collector, only!)

Description Volume (ml) lOO/Pack lOOO/Pack lOO/Pack(silanized)

Clear glass 2 5181-3375 5183-4491

Clear glass, write-on spot

2 5182-0543 5183-4492 5183-4494

Amber glass, write-on spot

2 5182-3376 5183-4493 5183-4495

Table 22 SnapTop Vials (Caps for Use with the Analytical Scale Fraction Collector, only!)


Clear glass 2 5182-0544 5183-4504 5183-4507


2 5182-0546 5183-4505 5183-4508


2 5182-0545 5183-4506 5183-4509

Table 23 Screw Top Vials (Caps for Use with the Analytical Scale Fraction Collector, only!)


Clear glass 2 5182-0714 5183-2067 5183-2070


6 Parts and MaterialsList of Recommended Vials and Caps


2 5182-0715 5183-2068 5183-2071


2 5182-0716 5183-2069 5183-2072

Table 24 Crimp Caps (Caps for Use with the Analytical Scale Fraction Collector, only!)

Description Septa 100/Pack

Silver aluminum Clear PTFE/red rubber 5181-1210

Silver aluminum Clear PTFE/red rubber 5183-4498 (1000/Pack)

Blue aluminum Clear PTFE/red rubber 5181-1215

Green aluminum Clear PTFE/red rubber 5181-1216

Red aluminum Clear PTFE/red rubber 5181-1217

Table 25 Snap Caps (Caps for Use with the Analytical Scale Fraction Collector, only!)


Clear polypropyIene Clear PTFE/red rubber 5182-0550

BIue polypropylene Clear PTFE/red rubber 5182-3458

Green polypropylene Clear PTFE/red rubber 5182-3457

Red polypropylene Clear PTFE/red rubber 5182-3459

Table 23 Screw Top Vials (Caps for Use with the Analytical Scale Fraction Collector, only!) (continued)



Parts and Materials 6List of Recommended Vials and Caps

Table 26 Screw Caps (Caps for Use with the Analytical Scale Fraction Collector, only!)


BIue polypropyIene Clear PTFE/red rubber 5182-0717

Green polypropyIene Clear PTFE/red rubber 5182-0718

Red polypropylene Clear PTFE/red rubber 5182-0719

BIue polypropylene Clear PTFE/silicone 5182-0720

Green polypropylene Clear PTFE/silicone 5182-0721

Red polypropyIene Clear PTFE/silicone 5182-0722


6 Parts and MaterialsList of Recommended Plates and Closing Mats

List of Recommended Plates and Closing Mats

Table 27 Recommended Plates and Closing Mats (Std. Well Plates and Closing Mats for Use with the Analytical Scale Fraction Collector, only!)

Description Volume (ml)

Package Part Number

96 polypropylene well-plate*

* For use with the analytical scale fraction collector, only!

0.5 10 5042-1386

96 polypropylene well-plate* 0.5 120 5042-1385

96 polypropylene deep well pate 1.0 50 5042-6454

96 polypropylene deep well-plate with glass inserts, caps and septa pre assembled*

0.35 1 5065-4402

glass inserts for 5065-4402* 0.35 1000 5188-5321

384 polypropylene well-plate* 0.1 30 5042-1388

96 polypropylene conical-well plate* 0.18 25 5042-8502

54 x 2ml vial plate* 1.5 6 G2255-68700

15 x 6ml vial plate 4.5 1 5022-6539

Plate for 27 self lock Eppendorf tubes* 0.5, 1.5, 2 1 5022-6538

Silicon Closing mats for 96 well-plate* 50 5042-1389


WARNING If you are using flammable solvents, remove the plates from the fraction collector after turning it 0FF. You avoid the risk of building explosive gas mixtures in the instrument.


Parts and Materials 6List of Recommended Plates and Closing Mats

WARNING If you are using flammable solvents, cover the plates. This action avoids the risk of building explosive gas mixtures in the instrument.

WARNING Closing mats with adhesive can give some contamination in the system. The adhesive is soluble in most of the solvents used in HPLC.

WARNING In general do not use closing mats with adhesive. The fraction collector has no prepunch needle, therefore the adhesive will clog the needle after several injections.


6 Parts and MaterialsTransport Unit Assembly (Preparative Scale)

Transport Unit Assembly (Preparative Scale)

Table 28 Transport Unit Assembly (Preparative Scale)


1 Transport Unit Assembly (Preparative Scale), includes items 2 and 4

G1364-60018

2a, b Tubing - diverter valve to needle, 0.8 mm ID G1364-86711

3 Diverter valve assembly G1364-61901

4 Needle carrier assembly preparative scale G1364-60011

5 Needle assembly preparative scale G1364-87201

6 Delay Calibration Adapter (not shown) G1364-87301

Figure 42 Transport Unit Assembly (Preparative Scale), partly displayed

2a

3

2b

4

5


Parts and Materials 6Transport Unit Assembly (Analytical Scale)

Transport Unit Assembly (Analytical Scale)

Table 29 Transport Unit Assembly (Analytical Scale)


1 Transport unit assembly (analytical scale), includes items 2 and 4

G1364-60019

2a, b Tubing - diverter valve to needle, 0.25 mm ID(order full kit)

G1364-68712

3 Diverter valve assembly G1364-61901

4 Needle carrier assembly analytical scale G1364-60022

5 Needle assembly analytical scale (50mm needle)Needle assembly semi-preparative scale (20mm needle)

G1367-87200G1364-87202

6 Wellplate adapter G1364-60021


6 Parts and MaterialsTransport Unit Assembly (Analytical Scale)

Figure 43 Transport Unit Assembly (Analytical Scale), partly displayed

2a

3

2b

4

5

6


Parts and Materials 6Needle Assemblies

Needle Assemblies

Table 30 Needle Assemblies


1 Needle assembly preparative scale G1364-87201

2 Needle assembly semi preparative scale (20 mm needle)For use in the analytical scale fraction collector at flow rates > 10 ml/min. or with high test tubes (48 mm - 75mm).

G1364-87202

3 Needle assembly analytical scale (50 mm needle) G1367-87200

Delay Calibration Adapter (not shown) G1364-87301

Figure 44 Needle Assemblies

1 2 3


6 Parts and MaterialsDiverter-Valve Assembly

Diverter-Valve Assembly

Table 31 Diverter-Valve Assembly (preparative and analytical scale)


1 Diverter-valve assembly G1364-61901

2 PIN screw 0515-1211

Figure 45 Diverter-Valve Assembly

2

1


Parts and Materials 6Tubing Kits

Tubing Kits

Table 32 Tubing Kits


1 Tubing kit preparative scale 0.8 mm ID (consists of items 2,and 3)

G1364-68711

2 Inlet/waste tubing assembly preparative scale 0.8 mm ID Order Item 1

3 Diverter valve to needle tubing preparative scale 0.8 mm ID Order Item 1

4 Tubing kit analytical scale 0.25 mm ID (consists of items 5,and 6)

G1364-68712

5 Inlet/waste tubing assembly analytical scale 0.25 mm ID Order Item 4

6 Injection valve to needle tubing analytical scale 0.25 mm ID Order Item 4

7 Low dispersion tubing kit analytical scale 0.15 mm IDfor flow rates < 1 ml/min

G1364-68723

8 Inlet/waste tubing assembly analytical scale 0.15 mm ID Order Item 7

9 injection valve to needle tubing analytical scale 0.15 mm ID Order Item 7

10 Finger tight fitting (pack of 2) 0100-1516

Figure 46 Tubing Kits

10

2, 5, 8

3, 6, 9


6 Parts and MaterialsInternal Tray Assembly

Internal Tray Assembly

Table 33 Internal Tray Assembly (Analytical and Preparative Scale)


1 Internal tray assembly analytical scale, includes items 2 – 7Internal tray assembly preparative scale, includes items 2 – 7

G1364-63114G1364-63113

2 Screw-seat-adapter 5022-2200

3 Seal funnel (reorder No., pack. of 10) G1364-68730

4a Waste Tubing Kit 0.5T (analytical scale) G1364-86708

4b Waste Tubing Kit 0.8T (preparative scale) G1364-86719

5 Funnel coupler G1364-43201

6 Tray internal Order Item 1

7 Flow delay sensor (not shown) Order Item 1

Figure 47 Internal Tray Assembly (Analytical Scale)

2

3

4 a,b

5

6


Parts and Materials 6Fraction Collector Accessory Kit

Fraction Collector Accessory Kit

Table 34 Fraction Collector Accessory Kit Contents G1364-68725


Wrench, open end, 4mm 1 8710-1534

Wrench, open end, 1/4-5/16 inch 1 8710-0510

Hex key 2.0 mm 1 8710-2476

Finger tight fittings, 1/16” f-120*

* Reorder gives pack of 2

3 0100-1516

Waste tubing (1.2 m)†

† Reorder gives 5 m

1 5062-2463

Y-Type Fitting for waste tubing‡

‡ Reorder gives pack of 10

1 5065-9971

CAN cable, 1 m 1 5181-1519

Air channel adapter 1 G1329-43200

Sticking clamp for corrugated waste tubing (large)**

** Reorder gives 10 clamps for corrugated waste tubing (large))

3 5065-9976

Sticking clamp for waste tubing (small)††

†† Reorder gives 10 clamps for waste tubing (small)

3 5065-9976

ESD wrist strap 1 9300-1408




6 Parts and MaterialsFraction Collector Accessory Kit



7Specifications

Performance Specifications for the Fraction Collectors 180


7 SpecificationsPerformance Specifications for the Fraction Collectors

Performance Specifications for the Fraction Collectors

Table 35 performance Specifications Agilent 1200 Series PREPARATIVE SCALE Fraction Collector (G1364B)

Type Specification

trigger modes Time slices, Peak (threshold, up- / downslope),Timetable (combination of time intervals and peak) andManual trigger (supported only with G1323B Control Module)Agilent 1200 Series DAD/MWD detectors (G1315A/B/C, G1365 A/B/C), the Agilent 1200 Series fluorescence detector and the Agilent G1946C/D, G1956A/B LC-MSD are fully supported other detectors can be used but are not supported for fraction collection.

operating modes Discrete fractions: default mode for all vessels. The flow is diverted to waste, while moving from one vessel position to the next vessel positionContinuous flow: optional, available only when using well plates. It is possible to move from one well plate position to the next one without diverting the flow into the well plate to waste

Fraction capacities and trays

4 x well-plates full tray (MTP)* (for use with deep well plates, only)2 × well-plates std. tray (MTP) (for use with deep well plates, only) + 10 × 2 ml vials* (+ 1 half tray)100 x 2 ml in std. tray (+ 1 half tray)*

3 x 40 x 2 ml in half tray*

3 x 15 x 6 ml in half tray*

Full tray with 40 test tubes (30 mm OD, max. height 100 mm, ~45 ml / tube)Full tray with 60 test tubes (25 mm OD, max. height 100 mm, ~25 ml / tube)Full tray with 126 test tubes (16 mm OD, max. height 100 mm, ~12 ml / tube)Full tray with 215 test tubes (12 mm OD, max. height 100 mm, ~7 ml / tube)Installed trays are automatically detected and identified. For the with uncapped vials, tests tubes and well plates, only!

test tube / plate sizes

Minimum 48 mm to 100 mm maximum

Maximum tube volume

ca. 45 ml


Specifications 7Performance Specifications for the Fraction Collectors

Maximum flow rate 100 ml / min (depending on viscosity and generated back pressure, max. 6 bar at the diverter valve)

Delay volumes [μl] Fraction collector inlet to diverter valve: ~500 (typical, depends on length of the tubing)Diverter valve: ~15Diverter valve to needle: ~110Needle: ~5

Delay calibration sensor

Single wavelength absorbance detector working at 654 nm, consisting of a LED and a photo diode

Diverter valve 3/2 Diverter valve with low internal volume (15 μl), switching time < 100 ms, maximum operating pressure 6 bar

cooling Optional (with additional G1330B), performance depending on ambient conditions and the volume of collected fractions

maximum capacity 3 fraction collectors in parallel plus one recovery fraction collector connected via 12-Position, 13-Port Selector valve (PN G1160A)

GLP features Early maintenance feedback (EMF), electronic records of maintenance and errors

Interfaces Controller-area network (CAN). optional; LAN or external contacts interfaceRS232C,APG-remote (for remote start / stop signals to / from other modules)Interface to G1330A ThermostatCAN-DC-out for operation of Agilentapproved external devices like valves

Safety features Leak detection and safe leak handling, error detection and display, exhaust fan for fume extraction of hazardous vapors

* Vials can be used as recommended by Agilent Technologies (see “List of Recommended Vials and Caps” on page 165 and “List of Recommended Plates and Closing Mats” on page 168) but must be uncapped. Only the 96 deep well-plates can be used (without closing mats, see “List of Recom-mended Plates and Closing Mats” on page 168)

Table 35 performance Specifications Agilent 1200 Series PREPARATIVE SCALE Fraction Collector (G1364B) (continued)

Type Specification



Table 36 Performance Specifications Agilent 1200 Series ANALYTICAL SCALE Fraction Collector (G1364C)

Type Specification

trigger modes Time slices, Peak (threshold, up- / downslope), Timetable (combination of time intervals and peak) and Manual trigger (supported only with G1323B Control Module)Agilent 1200 Series DAD/MWD detectors (G1315A/B/C, G1365 A/B/C), the Agilent 1200 Series fluorescence detector and the Agilent G1946C/D, G1956A/B LC-MSD are fully supported other detectors can be used but are not supported for fraction collection.

operating modes Discrete fractions: default mode for all vessels. The flow is diverted to waste, while moving from one vessel position to the next vessel positionContinuous flow: optional, available only when using the deep well plates. It is possible to move from one well plate position to the next one without diverting the flow into the well plate to wasteNeedle into location: Needle pushes into the vessel as deep as specified, for the use with capped vials and test tubes and well plates with closing matsDroplet setup mode: The tip of the fraction collector needle will initially move down to the bottom of the well. Then it will slowly move upwards while the fraction is collected. The droplet setup mode enables the fraction collector to collect small fractions without bubbles.



fraction vessel capacities and trays

• 4 x well-plates full tray (MTP)* • 2 x well-plates std. tray + 10 funnels with external containers* (+ 1

half tray)• 2 × well-plates std. tray (MTP) + 10 × 2 ml vials* (+ 1 half tray)• 100 x 2 ml in std. tray (+ 1 half tray)*

• 3 x 40 x 2 ml in half tray*

• 3 x 40 funnels in half tray• 3 x 15 x 6 ml in half tray*

• Full tray with 40 test tubes (30 mm OD, max. height 48 mm, ~20 ml vol.)

• Full tray with 60 test tubes (25 mm OD, max. height 48 mm)• Full tray with 126 test tubes (16 mm OD, max. height 48 mm)• Full tray with 215 test tubes (12 mm OD, max. height 48 mm)Installed trays are automatically detected and identified. Installed plates and vials can be detected when operating in the needle into location mode* max. height can be extended by using the short needle assembly G1364-87202

maximum tube / plate height

48 mm with long needle assembly G1367-87200 75 mm with short needle assembly G1364-87202

Maximum tube volume ca. 20 ml with 48 mm test tubes, ca. 30 ml with 75 mm test tubes or unlimited, if funnels are used with external containers.

Maximum flow rate 10 ml / min (depending on viscosity and generated back pressure, max. 6 bar at the diverter valve). The analytical scale fraction collector can be modified for flow rates > 10 ml/min.

delay volumes [μl] Fraction collector inlet to diverter valve: ~50 (typical, depends on the length of the tubing)Diverter valve: ~15Diverter valve to needle: ~10Needle: ~4

delay calibration sensor Single wavelength absorbance detector working at 654 nm, consisting of a LED and a photo diode

diverter valve 3/2 Diverter valve with low internal volume (15 μl), switching time < 100 ms, maximum operating pressure 6 bar

Table 36 Performance Specifications Agilent 1200 Series ANALYTICAL SCALE Fraction Collector (G1364C) (continued)

Type Specification



cooling Optional (with additional G1330B), performance depending on ambient conditions and the volume of collected fractions

maximum capacity 3 fraction collectors in parallel plus one recovery fraction collector connected via 12-Position, 13-Port Selector valve (PN G1160A)

GLP features Early maintenance feedback (EMF), electronic records of maintenance and errors

interfaces - Controller-area network (CAN). - optional; LAN or external contacts interface- RS232C,- APG-remote (for remote start / stop signals to / from other modules)- Interface to G1330A Thermostat- CAN-DC-out for operation of Agilent approved external devices like valves

safety features Leak detection and safe leak handling, error detection and display, exhaust fan for fume extraction of hazardous vapors

* Vials and well-plates and capped vials and well plates with closing mats can be used as recom-mended by Agilent Technologies (see “List of Recommended Vials and Caps” on page 165 and “List of Recommended Plates and Closing Mats” on page 168)

Table 36 Performance Specifications Agilent 1200 Series ANALYTICAL SCALE Fraction Collector (G1364C) (continued)

Type Specification




ASafety Information

Safety Information 188

Lithium Batteries Information 191

Radio Interference 192

Sound Emission 193

Solvent Information 194

Agilent Technologies on Internet 195


A Safety InformationSafety Information

Safety Information

The following general safety precautions must be observed during all phases of operation, service, and repair of this instrument. Failure to comply with these precautions or with specific warnings elsewhere in this manual violates safety standards of design, manufacture, and intended use of the instrument. Agilent Technologies assumes no liability for the customer’s failure to comply with these requirements.

General

This is a Safety Class I instrument (provided with terminal for protective earthing) and has been manufactured and tested according to international safety standards.

WARNING If you are using flammable solvents, remove the well-plates from the tray when you turn off the sampler. You avoid the risk of building explosive gas mixtures in the tray compartment.

WARNING If you are using flammable solvents, cover the well-plates with closing mats to avoid the risk of building explosive gas mixtures.

WARNING After a leak in the sampler, make sure the leak plane is cleaned and dry.


Safety Information ASafety Information

Operation

Before applying power, comply with the installation section. Additionally the following must be observed.

Do not remove instrument covers when operating. Before the instrument is switched on, all protective earth terminals, extension cords, auto-transformers, and devices connected to it must be connected to a protective earth via a ground socket. Any interruption of the protective earth grounding will cause a potential shock hazard that could result in serious personal injury. Whenever it is likely that the protection has been impaired, the instrument must be made inoperative and be secured against any intended operation.

Make sure that only fuses with the required rated current and of the specified type (normal blow, time delay, and so on) are used for replacement. The use of repaired fuses and the short-circuiting of fuseholders must be avoided.

Do not operate the instrument in the presence of flammable gases or fumes. Operation of any electrical instrument in such an environment constitutes a definite safety hazard.

Do not install substitute parts or make any unauthorized modification to the instrument.

Capacitors inside the instrument may still be charged, even though the instrument has been disconnected from its source of supply. Dangerous voltages, capable of causing serious personal injury, are present in this instrument. Use extreme caution when handling, testing and adjusting.

WARNING Any adjustment, maintenance, and repair of the opened instrument under voltage is forbidden.

WARNING Disconnect the instrument from the line and unplug the power cord before maintenance.


A Safety InformationSafety Information

Safety Symbols

Table 37 shows safety symbols used on the instrument and in the manuals.

Table 37 Safety Symbols

Symbol Description

The apparatus is marked with this symbol when the user should refer to the instruction manual in order to prevent risk of harm to the operator and to protect the apparatus against damage.

Indicates dangerous voltages.

Indicates a protected conductor terminal.

Eye damage may result from directly viewing the light produced by the Xenon flash lamp used in this product. Always turn the xenon flash lamp off before removing it.

!

WARNING A warning alerts you to situations that could cause physical injury or damage to the equipment. Do not proceed beyond a warning until you have fully understood and met the indicated conditions.

CAUTION A caution alerts you to situations that could cause a possible loss of data. Do not proceed beyond a caution until you have fully understood and met the indicated conditions.


Safety Information ALithium Batteries Information

Lithium Batteries Information

WARNING Danger of explosion if battery is incorrectly replaced. Replace only with the same or equivalent type recommended by the equipment manufacturer. Lithium batteries may not be disposed-off into the domestic waste.

Transportation of discharged Lithium batteries through carriers regulated by IATA/ICAO, ADR, RID, IMDG is not allowed. Discharged Lithium batteries shall be disposed off locally according to national waste disposal regulations for batteries.

WARNING Lithiumbatteri - Eksplosionsfare ved fejlagtic handtering. Udskiftning ma kun ske med batteri af samme fabrikat og type. Lever det brugte batteri tilbage til leverandoren.

WARNING Lithiumbatteri - Eksplosionsfare. Ved udskiftning benyttes kun batteri som anbefalt av apparatfabrikanten. Brukt batteri returneres appararleverandoren.

NOTE Bij dit apparaat zijn batterijen geleverd. Wanneer deze leeg zijn, moet u ze niet weggooien maar inleveren als KCA.


A Safety InformationRadio Interference

Radio Interference

Never use cables other than the ones supplied by Agilent Technologies to ensure proper functionality and compliance with safety or EMC regulations.

Test and Measurement

If test and measurement equipment is operated with equipment unscreened cables and/or used for measurements on open set-ups, the user has to assure that under operating conditions the radio interference limits are still met within the premises.


Safety Information ASound Emission

Sound Emission

Manufacturer’s Declaration

This statement is provided to comply with the requirements of the German Sound Emission Directive of 18 January 1991.

This product has a sound pressure emission (at the operator position) < 70 dB.

• Sound Pressure Lp < 70 dB (A)

• At Operator Position

• Normal Operation

• According to ISO 7779:1988/EN 27779/1991 (Type Test)


A Safety InformationSolvent Information

Solvent Information

Observe the following recommendations on the use of solvents.

Solvents

Brown glass ware can avoid growth of algae.

Always filter solvents, small particles can permanently block the capillaries. Avoid the use of the following steel-corrosive solvents:

• Solutions of alkali halides and their respective acids (for example, lithium iodide, potassium chloride, and so on).

• High concentrations of inorganic acids like nitric acid, sulfuric acid especially at higher temperatures (replace, if your chromatography method allows, by phosphoric acid or phosphate buffer which are less corrosive against stainless steel).

• Halogenated solvents or mixtures which form radicals and/or acids, for example:

2CHCl3 + O2 → 2COCl2 + 2HCl

This reaction, in which stainless steel probably acts as a catalyst, occurs quickly with dried chloroform if the drying process removes the stabilizing alcohol.

• Chromatographic grade ethers, which can contain peroxides (for example, THF, dioxane, di-isopropylether) such ethers should be filtered through dry aluminium oxide which adsorbs the peroxides.

• Solutions of organic acids (acetic acid, formic acid, and so on) in organic solvents. For example, a 1-% solution of acetic acid in methanol will attack steel.

WARNING This instrument should only be used with solvents that have an ignition temperature higher than 200oC!


Safety Information AAgilent Technologies on Internet

• Solutions containing strong complexing agents (for example, EDTA, ethylene diamine tetra-acetic acid).

• Mixtures of carbon tetrachloride with 2-propanol or THF.

Agilent Technologies on Internet

For the latest information on products and services visit our worldwide web site on the Internet at:

http://www.agilent.com

Select “Products” - “Chemical Analysis”

It will provide also the latest firmware of the Agilent 1200 Series modules for download.


A Safety InformationAgilent Technologies on Internet


Index

Aabove location, 48accessory kit, 177accessory kit contents, 18adjusting the tray height, 27Agilent on internet, 195air circulation, 13analytical needle assembly, 150application notes, 91arm length calibration, 135

Bbattery

safety information, 191bench space, 13

Ccable

connecting APG remote, 22, 24connecting CAN, 22, 24connecting GPIB, 22, 24connecting LAN, 22, 24connecting the ChemStation, 22, 24connecting the power, 22, 24

cable connections, 28capacities, 180, 184capillaries, 34changing the serial number, 124check-out procedure, 86ChemStation

configuration, 46cleaning the fraction collector, 138closing mats, 168collection mode, 48, 115collection order, 48

condensation, 14configuration, 46

collection order, 48fraction delay volumes, 48multiple fraction collectors, 50needle movement, 48trays, 46well plates, 49

Configure - Needle move, 117configuring well-plate types, 40contact control, 48control module

analysis view, 95changing the serial number, 124configure screens, 111control screens, 109detectors, 102EMF, 118error log, 120firmware update screens, 122fraction, 113fraction - delays, 113fraction - filling order, 114fractions, 106introduction to AFC views and

screens, 93maintenance log, 121manual trigger, 110method screens, 103recovery location, 115reserved locations, 114run times screen, 103sequence, 105serial number, 124settings screens, 97signal plot screens, 107status, 106system view, 109

tests, 117thermostat screen, 102timetable screens, 100trays, 112

cooling, 181, 185corrugated waste tubing, 26

Ddamaged packaging, 16data analysis, 78delay calibration sensor, 181, 184delay volume, 48

configuration, 48delay volumes, 181, 184delivery checklist, 16detectors, 68diverter valve, 181, 184diverter valve assembly, 174

Eelectrical connections

thermostat, 33EMF, 118environment, 12, 14error messages, 128, 131

Ffailure, 128fill levels

reset, 74fill volume

maximum, 71firmware update, 122flow connections, 34fraction, 113


Index

fraction - delays, 113fraction - filling order, 114fraction capacities and trays, 180fraction delay volumes, 48fraction list, 78fraction preview, 72fraction vessel capacities and trays, 184funnel, 158fuses, 12

GGLP features, 181, 185

Hhalf-tray combinations, 38height, 27home position, 132

Iinlet / waste tubings, 141inner capillary diameter, 83installing

corrugated waste tubing, 26trays, 36waste tubings, 36

installing the fraction collector, 25flow connections, 34interface cables, 25power cable, 25safety, 25

installing the thermostatted fraction collector

interface cables, 29power cable, 29preparation, 29safety, 25, 29

instrument status indicator, 130interfaces, 181, 185internal tray, 156, 176internet, 195into location, 48

Lliquid contact control, 48

Mmaintenance functions, 128, 131

home position, 132park arm position, 132parts exchange position, 131transport unit self alignment, 133

maximumcapacity, 181, 185flow rate, 181, 184height, 184sizes, 180tube volume, 180, 184

method, 49setup, 49, 67

missing parts, 16

NNeedle, 173needle, 148, 150needle assemblies, 173needle movement, 48, 85, 115needle type, 83

OOnline tick marks, 77operating modes, 180, 183optimization, 84

Ppark arm position, 132park transport unit assembly, 43parts and materials, 16, 161

accessory kit, 177closing mats, 168diverter valve assembly, 174internal tray, 176needle assemblies, 173plates, 168

recommended tubes, 164recommended vials and caps, 165sheet metal kit, 177test tubes, 164transport unit assembly, 170, 171trays, 162tubing kits, 175vials and caps, 165

parts exchange position, 131peak based, 68peak detectors, 68performance specifications, 179, 180

analytical scale, 183preparative scale, 180

physical specifications, 14fraction collector, 14thermostat, 15

plates, 168pooling, 80, 85power consideration, 12power cords, 13power requirements, 12power supply indicator, 129preparative needle assembly, 148preview, 72

Rrecommended tubes, 164recommended vials and caps, 165recovery, 81recovery fraction collector, 51recovery location, 115removing

tray compartment divider, 31repairing the fraction collector, 137repairs

analytical needle assembly, 150cleaning the fraction collector, 138funnel, 158inlet / waste tubings, 141internal tray, 156main procedures overview, 139needle, 148, 150


Index

preparative needle assembly, 148simple repairs, 138, 140valve to needle tubings, 145

report, 79reserved locations, 49, 114reset fill volumes, 74rinsing, 70, 85

Ssafety features, 185safety information

on lithium batteries, 191sample recovery, 47, 81semi-preparative operation, 82serial number, 124sheet metal kit, 177shipping, 43simple repairs, 138, 140site requirements, 12slope only, 69Specifications, 179specifications, 14, 179, 180stack configuration, 22, 24

rear view, 22, 24start location, 74start run, 76start sequence, 76status indicators, 128, 129storage, 14

Ttemperature, 14, 180test tube

height, 83test tubes, 83, 164tests, 117thermostat

electrical connections, 33threshold

upper, 69tick marks, 79time based, 70

timetable, 68, 70transport, 43transport unit assembly, 43, 170, 171

analytical scale, 171preparative scale, 170

transport unit self alignment, 128, 133transporting the fraction collector, 43tray combinations, 38tray compartment divider, 31tray height adjustment, 27trays, 46, 112, 162, 180, 184

installing, 36numbering of vessel positions, 38

trigger modes, 180, 183troubleshooting and test functions, 127tube height, 46tube volume, 46tubes, 164tubing kits, 175

UUIB, 69universal interface box, 69unpacking the fraction collector, 16upper threshold, 69

Vvalve to needle tubings, 145vial contents temperature, 180vials and caps, 165

Wwaste tubings, 36weight, 13well plates, 38, 49

configuration, 49working mode, 69


Index


© Agilent Technologies 2006Printed in Germany 02/06

*G1364-90010**G1364-90010*G1364-90010

www.agilent.com

Agilent Technologies

In This Book

This manual contains technical reference information about the Agilent 1200 Series fraction collectors. The manual describes the following:

• configuration and operation of the fraction collector,

• troubleshooting and test functions,

• simple repairs

• parts and materials,

• specifications,

• safety information.

agilent 1200 series analytical and preparative scale ... · pdf filea agilent 1200 series...

Documents